NO325228B1 - New mandelic acid derivatives and their use as well as pharmaceutical formulation - Google Patents

New mandelic acid derivatives and their use as well as pharmaceutical formulation Download PDF

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NO325228B1
NO325228B1 NO20032465A NO20032465A NO325228B1 NO 325228 B1 NO325228 B1 NO 325228B1 NO 20032465 A NO20032465 A NO 20032465A NO 20032465 A NO20032465 A NO 20032465A NO 325228 B1 NO325228 B1 NO 325228B1
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mmol
rotamer
nmr
mhz
aze
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NO20032465A
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NO20032465L (en
NO20032465D0 (en
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Tord Inghardt
Arne Svensson
Anders Johansson
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Astrazeneca Ab
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Priority claimed from SE0004458A external-priority patent/SE0004458D0/en
Priority claimed from SE0100965A external-priority patent/SE0100965D0/en
Priority claimed from SE0101239A external-priority patent/SE0101239D0/en
Priority claimed from SE0102921A external-priority patent/SE0102921D0/en
Application filed by Astrazeneca Ab filed Critical Astrazeneca Ab
Publication of NO20032465D0 publication Critical patent/NO20032465D0/en
Publication of NO20032465L publication Critical patent/NO20032465L/en
Publication of NO325228B1 publication Critical patent/NO325228B1/en

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Description

Foreliggende oppfinnelse angår mandelsyre-derivater og anvendelse derav samt farmasøytisk formulering. The present invention relates to mandelic acid derivatives and their use as well as pharmaceutical formulation.

Oppfinnelsens område Field of the invention

Foreliggende oppfinnelse angår følgelig nye farmasøytisk anvendelige forbindelser, spesielt forbindelser som er og/eller forbindelser som blir metabolisert til forbindelser som er, kompetitive inhibitorer av trypsin-lignende serinproteaser, spesielt trombin, anvendelse av dem som medikamenter, farmasøytiske preparater inneholdende dem og synteseveier for deres produksjon. The present invention therefore relates to new pharmaceutically usable compounds, in particular compounds which are and/or compounds which are metabolized to compounds which are, competitive inhibitors of trypsin-like serine proteases, in particular thrombin, their use as drugs, pharmaceutical preparations containing them and synthesis routes for their production.

Bakgrunn Background

Blodkoagulering er nøkkelprosessen involvert i både hemostase (dvs. forhindring av blodtap fra å skade kar) og trombose (dvs. dannelsen av en blodkoagel i et blodkar, som noen ganger fører til karobstruksjon). Blood coagulation is the key process involved in both hemostasis (i.e. preventing blood loss from damaging vessels) and thrombosis (i.e. the formation of a blood clot in a blood vessel, sometimes leading to vessel obstruction).

Koagulering er resultatet av en kompleks serie av enzymatiske reaksjoner. En av de ultimate trinnene i denne serien av reaksjoner er omdannelsen av proenzymet protrombin til det aktive enzymet trombin. Coagulation is the result of a complex series of enzymatic reactions. One of the ultimate steps in this series of reactions is the conversion of the proenzyme prothrombin to the active enzyme thrombin.

Trombin er kjent for å spille en sentral rolle i koagulering. Det aktiverer blodplater, hvilket fører til blodplateaggregering, omdanner fibrinogen til fibrin monomerer, som polymeriserer spontant til fibrin polymerer og aktiverer faktor XIII, som igjen kryssbinder polymerene for å danne uoppløselig fibrin. Videre, aktiverer trombin faktor V og faktor VIII hvilket fører til en "positive feedback" dannelse av trombin fra protrombin. Thrombin is known to play a central role in coagulation. It activates platelets, leading to platelet aggregation, converts fibrinogen to fibrin monomers, which polymerize spontaneously to fibrin polymers and activates factor XIII, which in turn cross-links the polymers to form insoluble fibrin. Furthermore, thrombin activates factor V and factor VIII which leads to a "positive feedback" formation of thrombin from prothrombin.

Ved å hemme aggregering av blodplater og dannelsen og kryssbinding av fibrin, vil effektive inhibitorer av trombin være forventet å vise antitrombotisk aktivitet. I tillegg, vil antitrombotisk aktivitet være forventet å være forbedret ved effektiv hemning av positiv feedback mekanisme. By inhibiting platelet aggregation and the formation and crosslinking of fibrin, effective inhibitors of thrombin would be expected to exhibit antithrombotic activity. In addition, antithrombotic activity would be expected to be enhanced by effective inhibition of the positive feedback mechanism.

Tidligere Teknikk Previous Technology

Tidlig utvikling av lav molekylvekt inhibitorer av trombin er beskrevet av Claesson i Blood Coagul. Fibrinol. (1994) 5,411. Early development of low molecular weight inhibitors of thrombin is described by Claesson in Blood Coagul. Fibrinol. (1994) 5,411.

Blombåck et al (i J. Clin. Lab. Invest. 24, suppl. 107, 59, (1969)) har beskrevet trombininhibitorer basert på aminosyresekvensen plassert rundt spaltningssetet for fibrinogen A kjeden. Av aminosyresekvensene beskrevet, har disse forfatterene foreslått at tripeptidetsekvensen Phe-Val-Arg (P9-P2-P1, nedenfor referert til som P3-P2-P1 sekvens) ville være den mest effektive inhibitor. Blombåck et al (in J. Clin. Lab. Invest. 24, suppl. 107, 59, (1969)) have described thrombin inhibitors based on the amino acid sequence located around the fibrinogen A chain cleavage site. Of the amino acid sequences described, these authors have proposed that the tripeptide sequence Phe-Val-Arg (P9-P2-P1, hereinafter referred to as the P3-P2-P1 sequence) would be the most effective inhibitor.

Trombininhibitorer basert på dipeptidyl-derivater med en I,a-aminoalkyl guanidin i Pl-stillingen er kjent fra US-patent nr 4,346,078 og internasjonal patentsøknad WO 93/11152. Lignende, strukturelt relaterte, dipeptidyl-derivater har også vært beskrevet. For eksempel beskriver internasjonal patentsøknad WO 94/29336 forbindelser med, for eksempel aminometylbenzamidiner, cykliske aminoalkylamidiner og cykliske aminoalkylguanidiner i Pl-stillingen (internasjonal patentsøknad WO 97/23499 beskriver prodroge av visse av disse forbindelser); europeisk patentsøknad 0 648 780, beskriver forbindelser med, for eksempel cykliske aminoalkylguanidiner i Pl-stilling. Thrombin inhibitors based on dipeptidyl derivatives with a I,a-aminoalkyl guanidine in the P1 position are known from US patent no. 4,346,078 and international patent application WO 93/11152. Similar, structurally related, dipeptidyl derivatives have also been described. For example, international patent application WO 94/29336 describes compounds with, for example, aminomethylbenzamidines, cyclic aminoalkylamidines and cyclic aminoalkylguanidines in the P1 position (international patent application WO 97/23499 describes prodrugs of certain of these compounds); European patent application 0 648 780 describes compounds with, for example, cyclic aminoalkylguanidines in the P1 position.

Trombininhibitorer basert på peptidyl-derivater som også har cykliske aminoalkylguanidiner (f.eks. enten 3- eller 4- aminometyl-l-amidino-piperidin) i Pl-stilling er kjent fra Europeisk Patentsøknader 0 468 231,0 559 046 og 0 641 779. Thrombin inhibitors based on peptidyl derivatives which also have cyclic aminoalkylguanidines (e.g. either 3- or 4-aminomethyl-1-amidino-piperidine) in the P1 position are known from European Patent Applications 0 468 231, 0 559 046 and 0 641 779 .

Trombininhibitorer basert på tripeptidyl-derivater med argininaldehyd i Pl-stillingen ble først beskrevet i europeisk patentsøknad 0 185 390. Thrombin inhibitors based on tripeptidyl derivatives with arginine aldehyde in the P1 position were first described in European patent application 0 185 390.

Mer nylig, er arginin aldehyd-baserte peptidyl-derivater, modifisert i P3-stilling, rapportert. For eksempel beskriver internasjonal patentsøknad WO 93/18060 hydroksysyrer, europeisk patentsøknad 0 526 877 des-aminosyrer og europeisk patentsøknad 0 542 525 O-metyl mandelsyrer i P3-stilling. More recently, arginine aldehyde-based peptidyl derivatives, modified in the P3 position, have been reported. For example, international patent application WO 93/18060 describes hydroxy acids, European patent application 0 526 877 des-amino acids and European patent application 0 542 525 O-methyl mandelic acids in the P3 position.

Inhibitorer av serinproteaser (f.eks. trombin) basert på elektrofile ketoner i Pl-stilling er også kjent. For eksempel beskriver europeisk patentsøknad 0 195 212 peptidyl I-ketoestere og amider, europeisk patentsøknad 0 362 002 fluoralkylamidketoner, europeisk patentsøknad 0 364 344I,#,A-triketoforbindelser og europeisk patentsøknad 0 530 167 I-alkoksyketonderivater av arginin i Pl-stilling. Inhibitors of serine proteases (eg thrombin) based on electrophilic ketones in the P1 position are also known. For example, European Patent Application 0 195 212 describes peptidyl I-ketoesters and amides, European Patent Application 0 362 002 fluoroalkylamide ketones, European Patent Application 0 364 344 I,#,A-triketo compounds and European Patent Application 0 530 167 I-Alkoxyketone derivatives of arginine in the P1 position.

Andre, strukturelt forskjellige, inhibitorer av trypsin-lignende serinproteaser basert på C-terminale boronsyrederivater av arginin og isotiouronium analoger derav er kjent fra europeisk patentsøknad 0 293 881. Other, structurally different, inhibitors of trypsin-like serine proteases based on C-terminal boronic acid derivatives of arginine and isothiouronium analogues thereof are known from European patent application 0 293 881.

Mer nylig, er trombininhibitorer basert på peptidyl-derivater beskrevet i europeisk patentsøknad 0 669 317 og internasjonal patentsøknader WO 95/35309, WO 95/23609, WO 96/25426, WO 97/02284, WO 97/46577, WO 96/32110, WO 96/31504, WO 96/03374, WO 98/06740, WO 97/49404, WO 98/57932, WO 99/29664, WO 00/35869 og WO 00/42059. More recently, thrombin inhibitors based on peptidyl derivatives are described in European Patent Application 0 669 317 and International Patent Applications WO 95/35309, WO 95/23609, WO 96/25426, WO 97/02284, WO 97/46577, WO 96/32110, WO 96/31504, WO 96/03374, WO 98/06740, WO 97/49404, WO 98/57932, WO 99/29664, WO 00/35869 and WO 00/42059.

Spesielt beskriver, WO 97/02284 og WO 00/42059 trombininhibitorer med substituerte mandelsyrer i P3 stilling. In particular, WO 97/02284 and WO 00/42059 describe thrombin inhibitors with substituted mandelic acids in the P3 position.

Imidlertid, er det fortsatt et behov for effektive inhibitorer av trypsin-lignende serinproteaser, så som trombin. Det er også et behov for forbindelser som har en fordelaktig farmakokinetisk profil og er selektive for å hemme trombin over andre serinproteaser, spesielt de involvert i hemostase. Forbindelser som viser kompetitiv hemmende aktivitet mot trombin ville være forventet å være spesielt anvendelige som antikoaguleirngsmidler og derfor i terapeutisk behandling av trombose og relaterte lidelser. However, there is still a need for effective inhibitors of trypsin-like serine proteases, such as thrombin. There is also a need for compounds that have an advantageous pharmacokinetic profile and are selective for inhibiting thrombin over other serine proteases, particularly those involved in hemostasis. Compounds exhibiting competitive inhibitory activity against thrombin would be expected to be particularly useful as anticoagulant agents and therefore in the therapeutic treatment of thrombosis and related disorders.

Beskrivelse ifølge foreliggende oppfinnelse Description according to the present invention

Ifølge oppfinnelsen tilveiebringes en forbindelse, kjennetegnet ved at den er valgt fra Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe); According to the invention, a compound is provided, characterized in that it is selected from Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe);

Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(OH) eller Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(OH) or

Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab, eller et farmasøytisk akseptabelt salt derav. Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab, or a pharmaceutically acceptable salt thereof.

Det er foretrukket at forbindelsen er kjennetegnet ved at den er Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(OMe) eller et farmasøytisk akseptebelt salt derav. It is preferred that the compound is characterized in that it is Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(OMe) or a pharmaceutically acceptable salt thereof.

Forbindelser med formlene XV og XX kan fremstilles ved omsetning av en tilsvarende forbindelse med formel X som ovenfor definert med, henholdsvis, en forbindelse med formel XVI som ovenfor definert eller en forbindelse med formel XXI, Compounds of the formulas XV and XX can be prepared by reacting a corresponding compound of the formula X as defined above with, respectively, a compound of the formula XVI as defined above or a compound of the formula XXI,

hvor R IL er som ovenfor definert, for eksempel under lignende betingelser som de beskrevet ovenfor for syntese av forbindelser med formel I. where R IL is as defined above, for example under similar conditions to those described above for the synthesis of compounds of formula I.

Forbindelser med formel XVI, XVIII, XIX, XDCA, XIXB, XIXC, XIXD og XXI er enten kommersielt tilgjengelig, er kjent i litteraturen eller kan oppnås enten analogt med prosessene beskrevet her eller ved konvensjonelle syntetiske prosedyrer, i henhold til standard teknikker, fra lett tilgjengelige utgangsmaterialer ved anvendelse av passende reagenser og reaksjonsbetingelser. For eksempel kan forbindelser med formlene XDCA og XDCB fremstilles ved omsetning av en tilsvarende forbindelse med formel XIXC eller XIXD (som passende) med en forbindelse med formel X, for eksempel under lignende betingelser som de beskrevet ovenfor. Compounds of formula XVI, XVIII, XIX, XDCA, XIXB, XIXC, XIXD and XXI are either commercially available, are known in the literature or can be obtained either analogously to the processes described herein or by conventional synthetic procedures, according to standard techniques, from easily available starting materials using appropriate reagents and reaction conditions. For example, compounds of formulas XDCA and XDCB can be prepared by reacting a corresponding compound of formula XIXC or XIXD (as appropriate) with a compound of formula X, for example, under similar conditions to those described above.

Forbindelser med formlene I og Ia, som definert ovenfor og derivater av disse, er referert til nedenfor som "forbindelsene ifølge foreliggende oppfinnelse". Compounds of formulas I and Ia, as defined above and derivatives thereof, are referred to below as "the compounds of the present invention".

Foretrukne forbindelser ifølge foreliggende oppfinnelse er for anvendelse som et farmasøytisk preparat. Preferred compounds according to the present invention are for use as a pharmaceutical preparation.

Forbindelsene ifølge foreliggende oppfinnelse kan vise tautomerisme. Alle tautomere former og blandinger derav omfattes av omfanget av foreliggende oppfinnelse. The compounds according to the present invention can show tautomerism. All tautomeric forms and mixtures thereof are covered by the scope of the present invention.

Forbindelser ifølge foreliggende oppfinnelse inneholder også to eller flere asymmetriske karbonatomer og kan derfor vise optisk og/eller diastereoisomerisme. Diastereoisomerer kan separeres ved anvendelse av konvensjonelle teknikker, f.eks. kromatografi. De forskjellige stereoisomerer kan isoleres ved separering av en racemisk eller annen blanding av forbindelsene ved anvendelse av konvensjonelle, f.eks. HPLC teknikker. Alternativt kan de ønskede optiske isomerer fremstilles ved omsetning av de passende optisk aktive utgangsmaterialer under betingelser som ikke vil forårsake racemisering eller epimerisering eller ved derivatisering, for eksempel med en homochiral syre fulgt av separering av diastereomerderivater ved konvensjonelle metoder (f.eks. HPLC, kromatografi over silika). Alle stereoisomerer omfattes av omfanget av foreliggende oppfinnelse. Compounds according to the present invention also contain two or more asymmetric carbon atoms and can therefore show optical and/or diastereoisomerism. Diastereoisomers can be separated using conventional techniques, e.g. chromatography. The different stereoisomers can be isolated by separating a racemic or other mixture of the compounds using conventional, e.g. HPLC techniques. Alternatively, the desired optical isomers can be prepared by reacting the appropriate optically active starting materials under conditions that will not cause racemization or epimerization or by derivatization, for example with a homochiral acid followed by separation of diastereomer derivatives by conventional methods (e.g. HPLC, chromatography over silica). All stereoisomers are covered by the scope of the present invention.

Forbindelser ifølge foreliggende oppfinnelse som kan nevnes omfatter Ph(3-Cl)(5-OCHF2)-CH(OH)C(0)-Aze-Pab (hvor Aze symboliserer azetidin-2-karboksylat (dvs. i ( R)-og/eller (5)-konformasjoner). Compounds according to the present invention which may be mentioned include Ph(3-Cl)(5-OCHF2)-CH(OH)C(0)-Aze-Pab (where Aze symbolizes azetidine-2-carboxylate (i.e. in ( R)- and /or (5) conformations).

Medisinsk og farmasøytisk anvendelse Medical and pharmaceutical application

Forbindelser ifølge foreliggende oppfinnelse kan i seg selv ha farmakologisk aktivitet. Compounds according to the present invention may in themselves have pharmacological activity.

I henhold til et ytterligere aspekt ved foreliggende oppfinnelse er det således gitt forbindelsene ifølge foreliggende oppfinnelse for anvendelse som farmasøytiske midler. According to a further aspect of the present invention, the compounds according to the present invention are thus provided for use as pharmaceutical agents.

Spesielt, er forbindelsene ifølge foreliggende oppfinnelse anvendelige ved at de er for anvendelse ved behandling av en tilstand hvor inhibisjon av trombin er nødvendig. In particular, the compounds of the present invention are useful in that they are for use in the treatment of a condition where inhibition of thrombin is required.

Forbindelsene ifølge foreliggende oppfinnelse er således forventet å være anvendelige i de tilstandene hvor hemning av trombin er nødvendig og/eller tilstander hvor antikoagulant terapi er angitt, omfattende de følgende: Behandling og/eller forebygging av trombose og hyperkoagulabilitet i blod og/eller vev til dyr inkludert mennesker. Det er kjent at hyperkoagulabilitet kan føre til thrombo-emboliske sykdommer. Lidelser forbundet med hyperkoagulabilitet og thrombo-emboliske sykdommer som kan nevnes omfatter arvet eller ervervet aktivert protein C resistens, så som faktor V-mutasjon (faktor V Leiden) og arvet eller ervervet mangler i antithrombin III, protein C, protein S, heparin kofaktor II. Andre tilstander kjent å være forbundet med hyperkoagulabilitet og thrombo-embolisk sykdom omfatter sirkulerende antifosfolipid antistoffer (Lupus antikoagulant), homocysteinemi, heparin fremkalt trombocytopeni og defekter i fibrinolyse, så vel som koaguleringssyndromer (f.eks. disseminert intravaskulær koagulering (DIC)) og vaskulær skade generelt (f.eks. på grunn av kirurgi). The compounds according to the present invention are thus expected to be applicable in the conditions where inhibition of thrombin is necessary and/or conditions where anticoagulant therapy is indicated, including the following: Treatment and/or prevention of thrombosis and hypercoagulability in blood and/or tissue of animals including humans. It is known that hypercoagulability can lead to thromboembolic diseases. Disorders associated with hypercoagulability and thromboembolic diseases that may be mentioned include inherited or acquired activated protein C resistance, such as factor V mutation (factor V Leiden) and inherited or acquired deficiencies in antithrombin III, protein C, protein S, heparin cofactor II . Other conditions known to be associated with hypercoagulability and thromboembolic disease include circulating antiphospholipid antibodies (Lupus anticoagulant), homocysteinemia, heparin-induced thrombocytopenia, and defects in fibrinolysis, as well as coagulation syndromes (eg, disseminated intravascular coagulation (DIC)) and vascular injury in general (eg due to surgery).

Behandling av tilstander hvor det er et uønskede overskudd av trombin uten tegn på hyperkoagulabilitet, for eksempel i neurodegenerative sykdommer så som Alzheimer' s sykdom. Treatment of conditions where there is an unwanted excess of thrombin without signs of hypercoagulability, for example in neurodegenerative diseases such as Alzheimer's disease.

Spesielle sykdomstilstander som kan nevnes omfatter terapeutisk og/eller profylaktisk behandling av venøs trombose (f.eks. DVT) og pulmonal emboli, arteriell trombose (f.eks. i myokardialt infarkt, ustabil angina, trombose-basert slag og perifer arteriell trombose) og systemisk emboli vanligvis fra atrium under atrial fibrillering (f.eks. ikke-valvulær atrial fibrillering) eller fra venstre hjertekammer etter transmural myokardialt infarkt eller forårsaket av kongestivt hjertesvikt; forebygging av re-okklusjon (dvs. trombose) etter thrombolyse, perkutan trans-luminal angioplasti (PTA) og koronar-bypass operasjoner; forhindring av re-trombose etter mikrokirurgi og vaskulær kirurgi generelt. Special disease states that may be mentioned include therapeutic and/or prophylactic treatment of venous thrombosis (eg DVT) and pulmonary embolism, arterial thrombosis (eg in myocardial infarction, unstable angina, thrombosis-based stroke and peripheral arterial thrombosis) and systemic embolism usually from the atrium during atrial fibrillation (eg, non-valvular atrial fibrillation) or from the left ventricle after transmural myocardial infarction or caused by congestive heart failure; prevention of re-occlusion (ie thrombosis) after thrombolysis, percutaneous trans-luminal angioplasty (PTA) and coronary bypass operations; prevention of re-thrombosis after microsurgery and vascular surgery in general.

Ytterligere indikasjoner omfatter terapeutisk og/eller profylaktisk behandling av disseminert intravaskulær koagulering forårsaket av bakterier, multippel traume, forgiftning eller hvilken som helst annen mekanisme; antikoagulant behandling når blodet er i kontakt med fremmede overflater i kroppen så som vaskulære podninger, vaskulære stenter, vaskulære katetere, mekaniske og biologiske protese ventiler eller hvilken som helst annen medisinsk anordning; og antikoagulant behandling når blodet er i kontakt med medisinske anordninger utenfor kroppen så som under kardiovaskulær kirurgi ved anvendelse av en hjerte-lunge maskin eller i haemodialyse; terapeutisk og/eller profylaktisk behandling av idiopatisk og voksen åndenødssyndrom, pulmonal fibrose etter behandling med stråling eller kjemoterapi, septisk sjokk, septikemi, inflammatoriske responser, som omfatter, men er ikke begrenset til, ødem, akutt eller kronisk aterosklerose så som koronar arteriell sykdom og dannelsen av aterosklerotiske plaque, cerebral arteriell sykdom, cerebralt infarkt, cerebral trombose, cerebral emboli, perifer arteriell sykdom, ischemi, angina (omfattende ustabil angina), reperfusjonsskade, restenose etter perkutan trans-luminal angioplasti (PTA) og koronar arterie bypass kirurgi. Additional indications include therapeutic and/or prophylactic treatment of disseminated intravascular coagulation caused by bacteria, multiple trauma, poisoning, or any other mechanism; anticoagulant treatment when the blood is in contact with foreign surfaces in the body such as vascular grafts, vascular stents, vascular catheters, mechanical and biological prosthetic valves or any other medical device; and anticoagulant treatment when the blood is in contact with medical devices outside the body such as during cardiovascular surgery using a heart-lung machine or in hemodialysis; therapeutic and/or prophylactic treatment of idiopathic and adult respiratory distress syndrome, pulmonary fibrosis after treatment with radiation or chemotherapy, septic shock, septicemia, inflammatory responses, which include, but are not limited to, edema, acute or chronic atherosclerosis such as coronary artery disease and the formation of atherosclerotic plaque, cerebral arterial disease, cerebral infarction, cerebral thrombosis, cerebral embolism, peripheral arterial disease, ischaemia, angina (comprehensive unstable angina), reperfusion injury, restenosis after percutaneous trans-luminal angioplasty (PTA) and coronary artery bypass surgery.

Forbindelser ifølge foreliggende oppfinnelse som hemmer trypsin og/eller trombin kan også være anvendelige ved behandling av pankreatitt. Compounds according to the present invention which inhibit trypsin and/or thrombin can also be useful in the treatment of pancreatitis.

Forbindelsene ifølge foreliggende oppfinnelse er således angitt både ved terapeutisk og/eller profylaktisk behandling av disse tilstander. The compounds according to the present invention are thus indicated both for therapeutic and/or prophylactic treatment of these conditions.

Forbindelsene ifølge foreliggende oppfinnelse vil normalt administreres oralt, intravenøst, subkutant, buckalt, rektalt, dermalt, nasalt, trakealt, bronkialt, ved hvilken som helst annen parenteral vei eller via inhalering, i form av farmasøytiske preparater omfattende forbindelsen ifølge foreliggende oppfinnelse enten som en fri base eller et farmasøytisk akseptabelt ikke-toksisk organisk eller uorganisk syreaddisjonssalt, i en farmasøytisk akseptable doseform. The compounds according to the present invention will normally be administered orally, intravenously, subcutaneously, buccally, rectally, dermally, nasally, tracheally, bronchially, by any other parenteral route or via inhalation, in the form of pharmaceutical preparations comprising the compound according to the present invention either as a free base or a pharmaceutically acceptable non-toxic organic or inorganic acid addition salt, in a pharmaceutically acceptable dosage form.

Foretrukne administreringsmetoder av forbindelser ifølge foreliggende oppfinnelse er oral. Foretrukne farmasøytiske preparater omfatter farmasøytiske preparater med modifisert frigjøring omfattende forbindelser ifølge foreliggende oppfinnelse. Betegnelsen farmasøytisk preparat med "modifisert frigjøring" vil være kjent for fagfolk å omfatte hvilket som helst preparat hvor begynnelsen og/eller rate av frigjøring av medikament (dvs. forbindelsen ifølge foreliggende oppfinnelse) er endret ved galeniske manipulasjoner og omfatter således definisjonen gitt i United States Pharmacopeia (USP XXII) på sidene xliii og xliv av innledende del, den relevante beskrivelsen i dokumentet inntas herved som referanse. Preferred methods of administration of compounds according to the present invention are oral. Preferred pharmaceutical preparations include pharmaceutical preparations with modified release comprising compounds according to the present invention. The term "modified release" pharmaceutical preparation will be known to those skilled in the art to include any preparation in which the onset and/or rate of drug release (ie, the compound of the present invention) has been altered by galenic manipulations and thus includes the definition given in the United States Pharmacopeia (USP XXII) on pages xliii and xliv of the introductory part, the relevant description in the document is hereby incorporated by reference.

Egnete modifiserte frigjøringspreparater kan således fremstilles av fagfolk i henhold til standardteknikker i farmasi (se for eksempel Pharmaceutisch Weekblad Vitenskapelig Edition, 6, 57 (1984); Medical Applications of Controlled Release, Vol II, eds. Langer og Wise (1984) Bocaraton, Florida, på sidene 1 til 34; Industriell Aspects ofpharmaceuticals, ed. Sandel, Swedish Pharmaceutical Press (1993) på sidene 93 til 104; og sidene 191 til 211 av " Pharmaceutics: The Science ofDosage Form Design", ed. M. E. Aulton (1988) Suitable modified release preparations can thus be prepared by those skilled in the art according to standard techniques in pharmacy (see, for example, Pharmaceutisch Weekblad Vitenskapelig Edition, 6, 57 (1984); Medical Applications of Controlled Release, Vol II, eds. Langer and Wise (1984) Bocaraton, Florida , at pages 1 to 34; Industrial Aspects of pharmaceuticals, ed. Sandel, Swedish Pharmaceutical Press (1993) at pages 93 to 104; and pages 191 to 211 of "Pharmaceutics: The Science of Dosage Form Design", ed. M. E. Aulton (1988)

(Churchill Livingston)). (Churchill Livingston)).

Foretrukete preparater med modifisert frigjøring omfatter således de hvor en hensiktsmessig forbindelse ifølge foreliggende oppfinnelse er innleiret i en polymer matriks. I dette henseende foretrekker vi at preparatene omfattende forbindelser ifølge foreliggende oppfinnelse blir tilveiebrakt for oral adrnimstrering i form av et såkalt "svelle" modifisert-firgjøringsystem eller en "geldannende matriks" modifisert-frigjøringssystem, hvor forbindelsen ifølge foreliggende oppfinnelse er gitt sammen med en polymer som sveller i et vandig medium (dvs. en "hydrofil gelldannende komponent"). Preferred preparations with modified release thus include those where a suitable compound according to the present invention is embedded in a polymer matrix. In this regard, we prefer that the preparations comprising compounds of the present invention are provided for oral administration in the form of a so-called "swelling" modified release system or a "gel-forming matrix" modified release system, where the compound of the present invention is provided together with a polymer which swells in an aqueous medium (ie a "hydrophilic gel-forming component").

Vi foretrekker spesielt at forbindelsene ifølge foreliggende oppfinnelse blir formulert sammen i et geleringsmatriks preparat omfattende iota-carrageenan og én eller flere nøytrale geleringspolymerer. We particularly prefer that the compounds according to the present invention are formulated together in a gelling matrix preparation comprising iota-carrageenan and one or more neutral gelling polymers.

Iota-carrageenan er fortrinnsvis til stede i slikt et foretrukket preparat i et nivå på mer enn 15 vekt%. Foretrukne kvaliteter av iota-carrageenan omfatter farmasøytisk kvalitet iota-carrageenan (tilgjengelig fra FMC Biopolymer), som har en viskositet på ikke mindre enn Iota-carrageenan is preferably present in such a preferred preparation at a level of more than 15% by weight. Preferred grades of iota-carrageenan include pharmaceutical grade iota-carrageenan (available from FMC Biopolymer), which has a viscosity of not less than

5 centipois (eps), fortrinnsvis i området 5-10 eps (for en 1,5% løsning oppvarmet til 82°C, etter som viskositeten blir målt ved 75°C med et Brookfield LV viscometer utstyrt med en #1 spindel drevet ved en hastighet på 30rpm) og teknisk kvalitet iota-carrageenan 5 centipois (eps), preferably in the range of 5-10 eps (for a 1.5% solution heated to 82°C, after which the viscosity is measured at 75°C with a Brookfield LV viscometer equipped with a #1 spindle driven at a speed of 30rpm) and technical quality iota-carrageenan

(tilgjengelig fra Fluka Biochemica), som fortrinnsvis har en viskositet på ikke mindre enn 14 mPa.s, for en 0,3 % vandig løsning oppvarmet til 20°C, etter hvor viskositeten blir målt ved anvendelse av et "fallingball" viscometer, av type Haake, anvendt sammen med en Lauda thermostat C3 og Hakke Mess-System III og ved anvendelse av gull-belagte rustfri stål baller med tetthet 7,8 g/cm<3>. (available from Fluka Biochemica), preferably having a viscosity of not less than 14 mPa.s, for a 0.3% aqueous solution heated to 20°C, after which the viscosity is measured using a "falling ball" viscometer, of type Haake, used together with a Lauda thermostat C3 and Hakke Mess-System III and using gold-coated stainless steel balls with a density of 7.8 g/cm<3>.

Nøytral geldannende polymer kan være en enkel eller en blanding av mer enn en, nøytral eroderbar polymer(er) som har geldannende egenskaper og som har hovedsakelig pH-uavhengig oppløselighet. Nøytral geleringspolymer er, fortrinnsvis, til stede i preparatet i et nivå på mer enn 10% men fortrinnsvis mer enn 20 vekt%. Neutral gel-forming polymer can be a single or a mixture of more than one, neutral erodible polymer(s) having gel-forming properties and having essentially pH-independent solubility. Neutral gelling polymer is preferably present in the composition at a level of more than 10% but preferably more than 20% by weight.

Egnete nøytrale geleringspolymerer omfatter polyetylenoksyd (PEO), derivater og medlemmer av PEO familien (for eksempel polyetylenglykol (PEG), fortrinnsvis eksisterende naturlig i fast stoff tilstand, med egnet molekylvekt eller viskositet). Hvis anvendt som en enkel nøytral geldannende polymer, har en PEO fortrinnsvis en MW på > 4 million (4M), svarende til en vandig løsning viskositetsområde på 1650-5500 mPa.s (eller 1650-5500 eps; målt for en 1% vandig løsning ved 25°C, ved anvendelse av et Brookfield RVF viscometer, med Nr. 2 spindel, ved 2 rpm). Andre eksempler på egnede PEOs omfatter en PEO med MW på rundt 5 million (5M), svarende til et vandig løsningsviskositetsområde på 5500 - 7500 mPa.s eller en PEO MW på rundt 8 millioner (8M), svarende til et vandig løsnings viskositetsområde på 10000-15000 mPa.s. Dette område dekker verdien for typisk løsningsviskositet (i eps) målt ved 25°C, angitt for denne polymeren, i USP 24/NF 19,2000 utgave, s.2285-2286. Hvis PEG blir anvendt som en enkel nøytral geldannende polymer har den fortrinnsvis en høy molekylvekt, for eksempel en MW på rundt 20000, svarende til et viskositetsområde på 2700-3500 mPa.s (eller 2700-3500 eps), målt ved anvendelse av en 50% vandig løsning (vekt/vekt) ved 20°C, ved anvendelse av et kapilærviscometer (Ubbelohde eller tilsvarende). [Ref: European pharmacopoeia 3rf Ed., 2000, Supplement, s. 908-909.] Suitable neutral gelling polymers include polyethylene oxide (PEO), derivatives and members of the PEO family (for example polyethylene glycol (PEG), preferably existing naturally in the solid state, with a suitable molecular weight or viscosity). If used as a single neutral gel-forming polymer, a PEO preferably has a MW of > 4 million (4M), corresponding to an aqueous solution viscosity range of 1650-5500 mPa.s (or 1650-5500 eps; measured for a 1% aqueous solution at 25°C, using a Brookfield RVF viscometer, with No. 2 spindle, at 2 rpm). Other examples of suitable PEOs include a PEO with a MW of around 5 million (5M), corresponding to an aqueous solution viscosity range of 5500 - 7500 mPa.s or a PEO MW of around 8 million (8M), corresponding to an aqueous solution viscosity range of 10,000 -15000 mPa.s. This range covers the value for typical solution viscosity (in eps) measured at 25°C, given for this polymer, in USP 24/NF 19,2000 edition, pp.2285-2286. If PEG is used as a simple neutral gel-forming polymer it preferably has a high molecular weight, for example a MW of around 20,000, corresponding to a viscosity range of 2700-3500 mPa.s (or 2700-3500 eps), measured using a 50 % aqueous solution (w/w) at 20°C, using a capillary viscometer (Ubbelohde or equivalent). [Ref: European pharmacopoeia 3rf Ed., 2000, Supplement, pp. 908-909.]

Andre egnete geldannende polymerer omfatter cellulosederivater så som hydroksypropylmetylcellulose (HPMC) eller hydroksyetylcellulose (HEC) med hensiktsmessige høye viskositeter (for eksempel "HPMC 10000 eps", "HPMC 15000 eps", "HEC type HH" eller "HEC type H"). Når anvendt som en enkel nøytral polymer, hydroksypropylmetylcellulose polymerer som "HPMC 10000 eps" og "HPMC 15000 eps" har, henholdsvis, tilsynelatende viskositeter på 7500-14000 mPa.s (eller 7500 - 14000 eps) og 11250-21000 mPa.s (eller 11250-21000 eps), når målt ved 20°C med en 2% (vekt/vekt) vandig løsning, beregnet med referanse til tørket substans, ved anvendelse av et kapilær viscometer (Ubbelohde eller ekvivalent). En type av hydroksyetylcellulosepolymer, for eksempel "Natrosol 250 Pharma, type HH", fra Hercules Incorporated (Aqualon), viser typisk en Brookfield viskositet på ca. 20,000 mPa.s ved anvendelse av en Brookfield Synchro-Lectric Modell LVF instrument, ved tilstander med 1% løsningskonsentrasjon, spindel nr. 4, spindel hastighet 30 rpm, faktor 200,25°C (Se Natrosol Physical and Chemical Properties booklet, 33.007-E6 (1993), p. 21). Other suitable gel-forming polymers include cellulose derivatives such as hydroxypropyl methylcellulose (HPMC) or hydroxyethyl cellulose (HEC) with suitably high viscosities (eg "HPMC 10000 eps", "HPMC 15000 eps", "HEC type HH" or "HEC type H"). When used as a simple neutral polymer, hydroxypropylmethylcellulose polymers such as "HPMC 10000 eps" and "HPMC 15000 eps" have, respectively, apparent viscosities of 7500-14000 mPa.s (or 7500 - 14000 eps) and 11250-21000 mPa.s ( or 11250-21000 eps), when measured at 20°C with a 2% (w/w) aqueous solution, calculated with reference to dried substance, using a capillary viscometer (Ubbelohde or equivalent). One type of hydroxyethyl cellulose polymer, for example "Natrosol 250 Pharma, type HH", from Hercules Incorporated (Aqualon), typically exhibits a Brookfield viscosity of about 20,000 mPa.s using a Brookfield Synchro-Lectric Model LVF instrument, at conditions of 1% solution concentration, spindle no. 4, spindle speed 30 rpm, factor 200.25°C (See Natrosol Physical and Chemical Properties booklet, 33.007- E6 (1993), p. 21).

Spesielle preparater som kan nevnes omfatter de hvor forbindelsen ifølge foreliggende oppfinnelse blir formulert sammen med iota-carageenan og HPMC (10,000 eps) i et 50:50 (vekt %) forhold eller sammen med iota-carageenan og PEO 4M i et 50:50 (vekt %) forhold. Special preparations that can be mentioned include those where the compound according to the present invention is formulated together with iota-carrageenan and HPMC (10,000 eps) in a 50:50 (wt%) ratio or together with iota-carrageenan and PEO 4M in a 50:50 ( weight %) ratio.

Foretrukne ytterligere tilsetningsmidler i slike preparater omfatter smøremidler, så som natriumstearylfumarat. Preferred additional additives in such preparations include lubricants, such as sodium stearyl fumarate.

Avhengig av lidelse og pasient som skal behandles og administreringsveien, kan preparatene administreres i varierende doser. Depending on the disorder and the patient to be treated and the route of administration, the preparations can be administered in varying doses.

Forbindelsene ifølge foreliggende oppfinnelse kan også bli kombinert og/eller ko-administrert med hvilket som helst antitrombotisk middel(er) med en forskjellig virkningsmekanisme, så som én eller flere av de følgende: antiblodplate midler acetylsalicylsyre, ticlopidin og clopidogrel; thromboksan reseptor og/eller syntetase inhibitorer; fibrinogen reseptorantagonister; prostacyclin etterligninger; fosfodiesterase inhibitorer; ADP-reseptor (P2T) antagonister; og inhibitorer av karboksypeptidase U The compounds of the present invention may also be combined and/or co-administered with any antithrombotic agent(s) with a different mechanism of action, such as one or more of the following: antiplatelet agents acetylsalicylic acid, ticlopidine and clopidogrel; thromboxane receptor and/or synthetase inhibitors; fibrinogen receptor antagonists; prostacyclin mimics; phosphodiesterase inhibitors; ADP receptor (P2T) antagonists; and carboxypeptidase U inhibitors

(CPU). (CPU).

Forbindelsene ifølge foreliggende oppfinnelse kan videre bli kombinert og/eller ko-administrert med trombolytiske midler så som én eller flere av vevsplasminogen activator (naturlig, rekombinant eller modifisert), streptokinase, urokinase, prourokinase, anisoylert plasminogen-streptokinase activator kompleks (APSAC), spyttkjertel plasminogen aktivatorer fra dyr og lignende, ved behandling av trombotiske sykdommer, spesielt myokardialt infarkt. The compounds according to the present invention can further be combined and/or co-administered with thrombolytic agents such as one or more of tissue plasminogen activator (natural, recombinant or modified), streptokinase, urokinase, prourokinase, anisoylated plasminogen-streptokinase activator complex (APSAC), salivary gland plasminogen activators from animals and the like, in the treatment of thrombotic diseases, especially myocardial infarction.

I henhold til et ytterligere aspekt ved foreliggende oppfinnelse er det således gitt et farmasøytisk preparat omfattende hvilke som helst av forbindelsene ifølge krav 1 eller et farmasøytisk akseptabelt salt derav, blandet sammen med en farmasytisk akseptabel adjuvant, fortynningsmiddel eller bærer. According to a further aspect of the present invention, there is thus provided a pharmaceutical preparation comprising any of the compounds according to claim 1 or a pharmaceutically acceptable salt thereof, mixed together with a pharmaceutically acceptable adjuvant, diluent or carrier.

Egnet daglig doser av forbindelsene ifølge foreliggende oppfinnelse i terapeutisk behandling av mennesker er ca. 0,001-100 mg/kg kroppsvekt ved peroral administrering og 0,001-50 mg/kg kroppsvekt ved parenteral administrering. Suitable daily doses of the compounds according to the present invention in the therapeutic treatment of humans are approx. 0.001-100 mg/kg body weight by oral administration and 0.001-50 mg/kg body weight by parenteral administration.

For å unngå tvil, som anvendt her, omfatter betegnelsen "behandling" terapeutisk og/eller profylaktisk behandling. For the avoidance of doubt, as used herein, the term "treatment" includes therapeutic and/or prophylactic treatment.

Foreliggende oppfinnelse vedrører videre anvendelse av en forbindelse ifølge krav 1 eller 2, som et aktivt ingrediens for fremstilling av et medikament for behandling av en tilstand hvor inhibisjon av trombin er nødvendig. Oppfinnelsen vedrører videre anvendelse av en forbindelse ifølge krav 1 eller 2, som et aktivt ingrediens for fremstilling av et medikament for behandling av en tilstand hvor antikoaguleringsterapi er indikert. Foreliggende oppfinnelse vedrører også anvendelse av en forbindelse ifølge krav 1 eller 2 som et aktivt ingrediens for fremstilling av et antikoaguleringsmiddel. The present invention further relates to the use of a compound according to claim 1 or 2, as an active ingredient for the production of a drug for the treatment of a condition where inhibition of thrombin is necessary. The invention further relates to the use of a compound according to claim 1 or 2, as an active ingredient for the preparation of a drug for the treatment of a condition where anticoagulation therapy is indicated. The present invention also relates to the use of a compound according to claim 1 or 2 as an active ingredient for the production of an anticoagulant.

Forbindelser ifølge foreliggende oppfinnelse har den fordelen at de kan være mer effektive, være mindre toksiske, være lenger virkende, har et bredere område med aktivitet, er kraftigere, produserer færre bivirkninger, blir lettere absorbert og/eller har en bedre farmakokinetisk profil (f.eks. høyere oral biotilgjengelighet og/eller lavere fjerning), og/eller har andre anvendelige farmakologiske, fysiske eller kjemiske, egenskaper i forhold til, forbindelser kjent i tidligere teknikk. Forbindelser ifølge foreliggende oppfinnelse kan ha ytterligere den fordel at de kan administreres sjeldnere enn forbindelser kjent i tidligere teknikk. Compounds according to the present invention have the advantage that they can be more effective, be less toxic, be longer acting, have a wider range of activity, are more potent, produce fewer side effects, are more easily absorbed and/or have a better pharmacokinetic profile (e.g. e.g. higher oral bioavailability and/or lower clearance), and/or have other applicable pharmacological, physical or chemical properties compared to compounds known in the prior art. Compounds according to the present invention may have the further advantage that they can be administered less frequently than compounds known in the prior art.

Biologiske Tester Biological Tests

De følgende testprosedyrer kan anvendes. The following test procedures can be used.

Test A Test A

Bestemmelse av Trombin Koaguleringstid ( TT) Determination of Thrombin Coagulation Time (TT)

Inhibitorløsningen (25 TL) blir inkubert med plasma (25 TL) i tre minutter. Human trombin (T 6769; Sigma Chem. Co eller Hematologic Teknologies) i bufferløsning, pH 7,4 (25 TL, 4,0 NIH enheter/ml), blir deretter tilsatt og størkningstiden målt i en automatisk anordning (KC 10; Amelunge). The inhibitor solution (25 TL) is incubated with plasma (25 TL) for three minutes. Human thrombin (T 6769; Sigma Chem. Co or Hematologic Technologies) in buffer solution, pH 7.4 (25 TL, 4.0 NIH units/ml), is then added and the clotting time measured in an automatic device (KC 10; Amelunge) .

Trombin-koaguleringstid (TT) blir uttrykt som absolutte verdier (sekunder) så vel som forholdet av TT uten inhibitor (TT0) til TT med inhibitor (TT;). Det sistnevnte forhold (område 1-0) blir plottet mot konsentrasjonen av inhibitor (log transformert) og tilpasset til sigmoidal dose-respons kurver i henhold til ligningen Thrombin clotting time (TT) is expressed as absolute values (seconds) as well as the ratio of TT without inhibitor (TT0) to TT with inhibitor (TT;). The latter ratio (range 1-0) is plotted against the concentration of inhibitor (log transformed) and fitted to sigmoidal dose-response curves according to the equation

y = a/[l+(x/IC50)<s>] y = a/[l+(x/IC50)<s>]

hvor: a = maksimum området, dvs. 1; s = stigning av dosen-respons kurven; og IC50 = konsentrasjonen av inhibitor som dobbler størkningstiden. Beregningene er prosessert på en PC ved anvendelse av programvare program GraFit Versjon 3, ved innstilling av where: a = maximum range, i.e. 1; s = slope of the dose-response curve; and IC50 = the concentration of inhibitor that doubles the clotting time. The calculations are processed on a PC using the software program GraFit Version 3, by setting

ligningen til: Start ved 0, definerer ende = 1 (Erithacus Programvare, Robin Leatherbarrow, Imperial College av Science, London, UK). the equation of: Start at 0, define end = 1 (Erithacus Software, Robin Leatherbarrow, Imperial College of Science, London, UK).

TestB TestB

Bestemmelse av Trombin Hemning med en Kromogen. Robot Forsøk Determination of Thrombin Inhibition with a Chromogen. Robot Experiment

Trombininhibitor potens blir målt med en kromogen substratmetode, i en Plato 3300 robot mikroplate processor (Rosys AG, CH-8634 Hombrechtikon, Switzerland), ved anvendelse av 96-brønn, halv volum mikrotiter-plater (Costar, Cambridge, MA, USA; Katt No 3690). Lagerløsninger av testsubstans i DMSO (72 |iL), 0,1-1 mmol/L, blir fortynnet i serie 1:3 (24 + 48 uL) med DMSO for å oppnå ti forskjellige konsentrasjoner, som blir analysert som prøver i forsøket. 2 |iL av testprøve blir fortynnet med 124 [iL forsøksbuffer, 12 jiL av kromogen substrat løsning (S-2366, Chromogenix, Molndal, Sweden) i forsøksbuffer og til slutt 12 (iL av I-trombinløsning (Human a-trombin, Sigma Kjemisk Co. eller Hematologic Teknologies) i forsøksbuffer, blir tilsatt og prøvene blandet. De endelige forsøkskonsentrasjoner er: testsubstans Thrombin inhibitor potency is measured with a chromogenic substrate method, in a Plato 3300 robotic microplate processor (Rosys AG, CH-8634 Hombrechtikon, Switzerland), using 96-well, half-volume microtiter plates (Costar, Cambridge, MA, USA; Katt No. 3690). Stock solutions of test substance in DMSO (72 µL), 0.1-1 mmol/L, are serially diluted 1:3 (24 + 48 µL) with DMSO to obtain ten different concentrations, which are analyzed as samples in the experiment. 2 µl of test sample is diluted with 124 µl of test buffer, 12 µl of chromogenic substrate solution (S-2366, Chromogenix, Molndal, Sweden) in test buffer and finally 12 µl of I-thrombin solution (Human a-thrombin, Sigma Kjemisk Co. or Hematologic Technologies) in test buffer, is added and the samples are mixed The final test concentrations are: test substance

0,00068 - 13,3 umol/L, S-2366 0,30 mmol/L, I-trombin 0,020 NIHU/ml. lineær absorbans del under 40 minutter inkubering ved 37°C blir anvendt for beregning av prosentdel hemning for testprøver, sammenlignet med blanke kontrollprøver uten inhibitor. ICso-robot verdi, svarende til inhibitorkonsentrasjonen som forårsaker 50% hemning av trombin aktivitet, blir beregnet fra en log konsentrasjon vs. % hemningskurve. 0.00068 - 13.3 umol/L, S-2366 0.30 mmol/L, I-thrombin 0.020 NIHU/ml. linear absorbance part during 40 minutes incubation at 37°C is used for calculation of percentage inhibition for test samples, compared to blank control samples without inhibitor. The IC 50 robot value, corresponding to the inhibitor concentration that causes 50% inhibition of thrombin activity, is calculated from a log concentration vs. % inhibition curve.

TestC TestC

Bestemmelse av hemningskonstant K for Human Trombin Determination of the inhibition constant K for Human Thrombin

Ki-bestemmelser blir gjort ved anvendelse av en kromogen substrat metode, utført ved 37°C på en Cobas Bio centrifugal analysator (Roche, Basel, Switzerland). Gjenværende enzymaktivitet etter inkubering av human I-trombin med forskjellige konsentrasjoner av testforbindelse blir bestemt ved tre forskjellige substrat konsentrasjoner og blir målt som forandringen i optisk absorbans ved 405 nm. Ki determinations are made using a chromogenic substrate method, performed at 37°C on a Cobas Bio centrifugal analyzer (Roche, Basel, Switzerland). Residual enzyme activity after incubation of human I-thrombin with different concentrations of test compound is determined at three different substrate concentrations and is measured as the change in optical absorbance at 405 nm.

Testforbindelseløsninger (100 TL; normalt i buffer eller saltvann inneholdende BSA 10 g/L) blir blandet med 200 TL av human I-trombin (Sigma Kjemisk Co) i forsøksbuffer Test compound solutions (100 TL; normally in buffer or saline containing BSA 10 g/L) are mixed with 200 TL of human I-thrombin (Sigma Chemical Co) in test buffer

(0,05 mol/L Tris-HCl pH 7,4, ionestyrke 0,15 regulert med NaCl) inneholdende BSA (10 g/L) og analysert som prøver i Cobas Bio. En 60 TL prøve, sammen med 20 TL vann, blir satt til 320 TL av substratet S-2238 (Chromogenix AB, Molndal, Sverige) i forsøksbuffer og absorbanseforandringen (AA/min) blir overvåket. De endelige konsentrasjoner av S-2238 er 16,24 og 50 pmol/L og av trombin 0,125 NIH U/ml. (0.05 mol/L Tris-HCl pH 7.4, ionic strength 0.15 adjusted with NaCl) containing BSA (10 g/L) and analyzed as samples in Cobas Bio. A 60 TL sample, together with 20 TL water, is added to 320 TL of the substrate S-2238 (Chromogenix AB, Molndal, Sweden) in test buffer and the absorbance change (AA/min) is monitored. The final concentrations of S-2238 are 16.24 and 50 pmol/L and of thrombin 0.125 NIH U/ml.

Lineær-rate reaksjonsrate blir anvendt for å konstruere Dixon plott, dvs. diagrammer av inhibitorkonsentrasjon vs. l(AA/min). For reversible, kompetitive inhibitorer, datapunktene for de forskjellige substratkonsentrasjoner danner typisk lineære linjer som krysses ved x = -K,. Linear-rate reaction rates are used to construct Dixon plots, i.e. diagrams of inhibitor concentration vs. l(AA/min). For reversible, competitive inhibitors, the data points for the various substrate concentrations typically form linear lines that intersect at x = -K,.

TestD TestD

Bestemmelse av Aktivert Partiell Thromboplastin Tid ( APTT) Determination of Activated Partial Thromboplastin Time (APTT)

APTT blir bestemt i samlet normalt humant citrert plasma med reagenset PTT Automatisert 5 fremstilt av Stago. Inhibitorene blir satt til plasmaet (10 TL inhibitorløsning til 90 TL plasma) og inkubert med APTT reagens i 3 minutter fulgt av tilsetning av 100 TL av kalsiumklorid-løsning (0,025 M) og APTT blir bestemt ved anvendelse av koaguleringsanalyser KC10 (Amelunge) i henhold til instruksjoner av produsent produsenten. APTT is determined in pooled normal human citrated plasma with the reagent PTT Automated 5 manufactured by Stago. The inhibitors are added to the plasma (10 TL inhibitor solution to 90 TL plasma) and incubated with APTT reagent for 3 minutes followed by the addition of 100 TL of calcium chloride solution (0.025 M) and APTT is determined using coagulation assays KC10 (Amelunge) according to to the instructions of the manufacturer the manufacturer.

Størkningstiden blir uttrykt som absolutte verdier (sekunder) så vel som forholdet av APTT uten inhibitor (APTT0) til APTT med inhibitor (APTTi). Det sistnevnte forhold (område 1-0) blir plottet mot konsentrasjonen av inhibitor (log transformert) og tilpasset til sigmoidale dose-respons kurver i henhold til ligningen Clotting time is expressed as absolute values (seconds) as well as the ratio of APTT without inhibitor (APTT0) to APTT with inhibitor (APTTi). The latter ratio (range 1-0) is plotted against the concentration of inhibitor (log transformed) and fitted to sigmoidal dose-response curves according to the equation

y = a/[l+(x/IC50)<s>] y = a/[l+(x/IC50)<s>]

hvor: a = maksimumområde, dvs. 1; s = stigning av dosen-respons kurven; og IC50 = konsentrasjonen av inhibitor som dobbler størkningstiden. Beregningene er prosessert på en PC ved anvendelse av programvare program GraFit Versjon 3, ved innstilling av ligning til: Start ved 0, definerer ende = 1 (Erithacus Programvare, Robin Leatherbarrow, Imperial College av Science, London, UK). where: a = maximum range, i.e. 1; s = slope of the dose-response curve; and IC50 = the concentration of inhibitor that doubles the clotting time. The calculations are processed on a PC using the software program GraFit Version 3, by setting the equation to: Start at 0, define end = 1 (Erithacus Software, Robin Leatherbarrow, Imperial College of Science, London, UK).

IC50APTT er definert som konsentrasjonen av inhibitor i humant plasma som dobblet den aktiverte Partielle Thromboplastin Tiden. IC50APTT is defined as the concentration of inhibitor in human plasma that doubled the activated Partial Thromboplastin Time.

TestE TestE

Bestemmelse av Trombintid ex vivo Determination of Thrombin Time ex vivo

Hemning av trombin etter oral eller parenteral administrering av forbindelsene ifølge foreliggende oppfinnelse, oppløst i etanol:SolutolK:vann (5:5:90), blir undersøkt i bevisste rotter som, én eller to dager før forsøket, blir utstyrt med et kateter for blodprøvetakning fra carotid arterien. På eksperimentdagen blir blodprøver tatt ved bestemte ganger etter administreringen av forbindelsen til plastrør inneholdende 1 del natriumcitratløsning (0,13 mol pr. L) og 9 deler blod. Rørene blir sentrifugert for å oppnå blodplate manglende plasma. 50 uL av plasmaprøver blir utfelt med 100 uL kald acetonitril. Prøvene blir sentrifugert i 10 minutter ved 4000 rpm. 75 uL av supernatanten blir fortynnet med 75 uL 0,2% maursyre. 10 uL volum av de resulterende løsninger blir analysert ved LC-MS/MS og konsentrasjonene av trombininhibitor blir bestemt ved anvendelse av standardkurver. Inhibition of thrombin after oral or parenteral administration of the compounds of the present invention, dissolved in ethanol:SolutolK:water (5:5:90), is investigated in conscious rats which, one or two days before the experiment, are fitted with a catheter for blood sampling from the carotid artery. On the day of the experiment, blood samples are taken at specific times after the administration of the compound into plastic tubes containing 1 part sodium citrate solution (0.13 mol per L) and 9 parts blood. The tubes are centrifuged to obtain platelet-deficient plasma. 50 uL of plasma samples are precipitated with 100 uL of cold acetonitrile. The samples are centrifuged for 10 minutes at 4000 rpm. 75 µL of the supernatant is diluted with 75 µL of 0.2% formic acid. 10 µL volumes of the resulting solutions are analyzed by LC-MS/MS and the concentrations of thrombin inhibitor are determined using standard curves.

Test F Test F

Bestemmelse av Plasma Fjerning i Rotte Determination of Plasma Removal in the Rat

Plasma fjerning blir beregnet i sprague dawley hannrotter. Forbindelsen blir oppløst i vann og administrert som en subkutan bolus injeksjon i en dose på 4 fimol/kg. Blodprøver blir oppsamlet ved hyppige intervaller opptil 5 timer etter medikamentadministrering. Blodprøver blir sentrifugert og plasma blir separert fra blodceller og overført til medisinglass inneholdende citrat (10% endelig konsentrasjon). 50 uL av plasmaprøver blir utfelt med 100 uL kald acetonitril. Prøvene blir sentrifugert i 10 minutter ved 4000 rpm. Plasma clearance is calculated in male Sprague Dawley rats. The compound is dissolved in water and administered as a subcutaneous bolus injection at a dose of 4 fimol/kg. Blood samples are collected at frequent intervals up to 5 hours after drug administration. Blood samples are centrifuged and plasma is separated from blood cells and transferred to vials containing citrate (10% final concentration). 50 uL of plasma samples are precipitated with 100 uL of cold acetonitrile. The samples are centrifuged for 10 minutes at 4000 rpm.

75 uL av supernatanten blir fortynnet med 75 uL av 0,2% maursyre. 10 uL volum av de resulterende løsninger blir analysert ved LC-MS/MS og konsentrasjonene av trombininhibitor blir bestemt ved anvendelse av standardkurver. Området under plasmakonsentrasjon-tid profilen blir beregnet ved anvendelse av log/lineær trapezoidal regel og ekstrapolert til uendelig tid. Plasma fjerning(CL) av forbindelsen blir deretter bestemt som 75 µL of the supernatant is diluted with 75 µL of 0.2% formic acid. 10 µL volumes of the resulting solutions are analyzed by LC-MS/MS and the concentrations of thrombin inhibitor are determined using standard curves. The area under the plasma concentration-time profile is calculated using the log/linear trapezoidal rule and extrapolated to infinite time. Plasma clearance (CL) of the compound is then determined as

CL=Dose/AUC CL=Dose/AUC

Verdiene er angitt i ml/min/kg. The values are given in ml/min/kg.

TestG TestG

Bestemmelse av in vitro Stabilitet Determination of in vitro Stability

Levermikrosomer blir fremstilt fra Sprague-Dawley rotter og humane leverprøver i henhold til indre SOP. Forbindelsene blir inkubert ved 37°C ved en total mikrosom-protein konsentrasjon på 3 mg/ml i en 0,05 mol/L TRIS buffer ved pH 7,4, i nærvær av kofaktorer NADH (2,5 mmol/L) og NADPH (0,8 mmol/L). De innledende konsentrasjonene av forbindelse er 5 eller 10 u mol/L. Prøver blir tatt for analyse opptil 60 minutter etter starten av inkuberingen. Den enzymatiske aktiviteten i oppsamlet prøve blir umiddelbart stanset ved tilsetning av 20% myristinsyre ved et volum svarende til 3,3% av totalt prøvevolum. Konsentrasjonen av forbindelse gjenværende (ENDELIG KONS.) i 60 min. prøve blir bestemt ved hjelp av LCMS ved anvendelse av en prøve oppsamlet ved null tid som referanse (START KONS.). % av nedbrutt trombininhibitor blir beregnet som: Liver microsomes are prepared from Sprague-Dawley rat and human liver samples according to internal SOP. The compounds are incubated at 37°C at a total microsome protein concentration of 3 mg/ml in a 0.05 mol/L TRIS buffer at pH 7.4, in the presence of cofactors NADH (2.5 mmol/L) and NADPH (0.8 mmol/L). The initial concentrations of compound are 5 or 10 µmol/L. Samples are taken for analysis up to 60 minutes after the start of incubation. The enzymatic activity in the collected sample is immediately stopped by the addition of 20% myristic acid at a volume corresponding to 3.3% of the total sample volume. The concentration of compound remaining (FINAL CONC.) in 60 min. sample is determined by LCMS using a sample collected at zero time as a reference (START CONC.). % of degraded thrombin inhibitor is calculated as:

TestH TestH

Arteriell Trombose Modell Arterial Thrombosis Model

Karskade blir fremkalt ved påføring av jern(III) klorid (FeCla) topisk til carotid arterien. Rotter blir bedøvet med en intraperitoneal injeksjon av natriumpentobarbital (80 mg/kg; Apoteksbolaget; Umeå, Sweden), fulgt av kontinuerlig infusjon (12 mg/kg/t) gjennom hele forsøket. Rottekroppstemperaturen blir holdt ved 38°C gjennom hele forsøket ved ytre oppvarmning. Forsøket begynner med en 5 minutter lang kontrollperiode. Fem minutter senere, blir human I-fibrinogen (80 kBq; IM53; Amersham Internasjonal, Buckinghamshire, UK) gitt intravenøst og blir anvendt som en markør for den påfølgende innføringen av fibrin(ogen) til trombe. Proximal ende av karotidarteriesegmentet blir plassert i et plastrør (6 mm; Silastic®; Dow Corning, MI, USA) åpnet i lengderetningen, inneholdende FeCl3-fuktet (2 uL; 55% vekt/vekt; Merck, Darmstadt, Tyskland) filterpapir (diameter 3 mm; 1F; Munktell, Grycksbo, Sweden). Venstre karotidarterie blir eksponert for FeCl3 i 10 minutter og blir deretter fjernet fra plastrørene og fuktet i saltvann. Femti minutter senere, blir karotidarterien fjernet og skyllet i saltvann. Referanseblodprøver blir også tatt for bestemmelse av blod <125>I-aktivitet, 10 minutter etter injeksjon av <1>25I-fibrinogen og ved slutten av forsøket. <19>SI-aktiviteten i referanseblodprøver og karsegmentet blir målt i en gammateller (1282 Compugamma; LKB Wallac Oy, Turku, Finland) på samme dag som forsøket blir utført. Trombestørrelsen blir bestemt som mengden av I-aktivitet inkorporert i karsegment i relasjon til I-aktiviteten i blodet (cpm/mg). Vascular damage is induced by applying iron(III) chloride (FeCla) topically to the carotid artery. Rats are anesthetized with an intraperitoneal injection of sodium pentobarbital (80 mg/kg; Apoteksbolaget; Umeå, Sweden), followed by continuous infusion (12 mg/kg/h) throughout the experiment. The rat's body temperature is maintained at 38°C throughout the experiment by external heating. The experiment begins with a 5-minute long control period. Five minutes later, human I-fibrinogen (80 kBq; IM53; Amersham International, Buckinghamshire, UK) is given intravenously and is used as a marker for the subsequent incorporation of fibrin(ogen) into thrombus. The proximal end of the carotid artery segment is placed in a plastic tube (6 mm; Silastic®; Dow Corning, MI, USA) opened longitudinally, containing FeCl3-moistened (2 µL; 55% w/w; Merck, Darmstadt, Germany) filter paper (diameter 3 mm; 1F; Munktell, Grycksbo, Sweden). The left carotid artery is exposed to FeCl3 for 10 minutes and is then removed from the plastic tubes and moistened in saline. Fifty minutes later, the carotid artery is removed and flushed in saline. Reference blood samples are also taken for determination of blood <125>I activity, 10 minutes after injection of <1>25I-fibrinogen and at the end of the experiment. <19>SI activity in reference blood samples and the vessel segment is measured in a gamma counter (1282 Compugamma; LKB Wallac Oy, Turku, Finland) on the same day that the experiment is performed. The thrombus size is determined as the amount of I activity incorporated into the vessel segment in relation to the I activity in the blood (cpm/mg).

Generelle Eksperimentelle Detaljer General Experimental Details

TLC ble utført på silikagel. Chiral HPLC analyse ble utført ved anvendelse av en 46 mm X 250 mm Chiralcel OD kolonne med en 5 cm guard kolonne. Kolonnetemperaturen ble holdt ved 35°C. En strømningshastighet på 1,0 ml/min ble anvendt. En Gilson 115 UV detektor ved 228 nm ble anvendt. Den mobile fasen besto av heksaner, etanol og trifluroeddiksyre og det passende forhold er listet opp for hver forbindelse. Typisk, ble produktet oppløst i en minimal mengde av etanol og dette ble fortynnet med den mobile fasen. TLC was performed on silica gel. Chiral HPLC analysis was performed using a 46 mm X 250 mm Chiralcel OD column with a 5 cm guard column. The column temperature was maintained at 35°C. A flow rate of 1.0 ml/min was used. A Gilson 115 UV detector at 228 nm was used. The mobile phase consisted of hexanes, ethanol and trifluoroacetic acid and the appropriate ratio is listed for each compound. Typically, the product was dissolved in a minimal amount of ethanol and this was diluted with the mobile phase.

LC-MS/MS ble utført ved anvendelse av et HP-1100 instrument utstyrt med en CTC-PAL injector og en 5 um, 4x100 mm ThermoQuest, Hypersil BDS-C18 kolonne. En API-3000 (Sciex) MS detektor ble anvendt. Strømningshastigheten var 1,2 ml/min og den mobile fasen (gradient) besto av 10-90% acetonitril med 90-10% av 4 mM vandig ammoniumacetat, begge inneholdende 0,2% maursyre. LC-MS/MS was performed using an HP-1100 instrument equipped with a CTC-PAL injector and a 5 µm, 4x100 mm ThermoQuest, Hypersil BDS-C18 column. An API-3000 (Sciex) MS detector was used. The flow rate was 1.2 ml/min and the mobile phase (gradient) consisted of 10-90% acetonitrile with 90-10% of 4 mM aqueous ammonium acetate, both containing 0.2% formic acid.

<*>H NMR spektra ble registrert ved anvendelse av tetrametylsilan som den indre standard. <13>c NMR spektra ble registrert ved anvendelse av oppførte deutererte løsningsmidler som den indre standard. <*>H NMR spectra were recorded using tetramethylsilane as the internal standard. <13>c NMR spectra were recorded using listed deuterated solvents as the internal standard.

Eksempel 1 Example 1

Ph( 3- Cl¥5- OCHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( OcBu) Ph( 3- Cl¥5- OCHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( OcBu)

(i) 3- klor- 5- metoksvbenzaldehvd (i) 3- chloro- 5- methoxybenzaldehyde eq

3,5-dikloranisol (74,0 g, 419 mmol) i THF (200 ml) ble satt dråpevis til magnesiummetall (14,2 g, 585 mmol, pre-vasket med 0,5 N HC1) i THF (100 ml) ved 25°C. Etter tilsetningen, ble 1,2-dibrometan 3,5-Dichloroanisole (74.0 g, 419 mmol) in THF (200 mL) was added dropwise to magnesium metal (14.2 g, 585 mmol, pre-washed with 0.5 N HCl) in THF (100 mL) at 25°C. After the addition, 1,2-dibromoethane

(3,9 g, 20,8 mmol) tilsatt dråpevis. Den resulterende mørke brune blanding ble oppvarmet ved tilbakeløp i 3 timer. Blandingen ble avkjølt til 0°C og MN-dimetylformamid (60 ml) ble tilsatt i én porsjon. Blandingen ble fordelt med dietyleter (3 x 400 ml) og 6N HC1 (500 ml). De samlede organiske ekstrakter ble vasket med saltvann (300 ml), tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir en olje. Flash kromatografi (2x) på silikagel (3.9 g, 20.8 mmol) added dropwise. The resulting dark brown mixture was heated at reflux for 3 hours. The mixture was cooled to 0°C and MN-dimethylformamide (60 mL) was added in one portion. The mixture was partitioned with diethyl ether (3 x 400 mL) and 6N HCl (500 mL). The combined organic extracts were washed with brine (300 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give an oil. Flash chromatography (2x) on silica gel

under eluering med Heks:EtOAc (4:1) ga sub-tittelforbindelsen (38,9 g, 54%) som en gul olje. eluting with Hex:EtOAc (4:1) gave the sub-title compound (38.9 g, 54%) as a yellow oil.

'H NMR (300 MHz, CDC13) A 9,90 (s, 1H), 7,53 (s, 1H), 7,38 (s, 1H), 7,15 (s, 1H), 3,87 (s, 3H). 1 H NMR (300 MHz, CDCl 3 ) Δ 9.90 (s, 1H), 7.53 (s, 1H), 7.38 (s, 1H), 7.15 (s, 1H), 3.87 ( pp, 3H).

(ii) 3 - klor- 5- hvdroksybenzaldehvd (ii) 3 - chloro- 5- hydroxybenzaldehyde eq

En løsning av 3-klor-5-metoksybenzaldehyd (22,8 g, 134 mmol; se trinn (i) ovenfor) i CH2C12 (250 ml) ble avkjølt til 0°C. Bortribromid (15,8 ml, 167 mmol) ble tilsatt dråpevis over 15 min. Etter omrøring, av reaksjonsblandingen i 2 timer, ble H2O (50 ml) tilsatt langsomt. Løsningen ble deretter ekstrahert med Et20 (2 x 100 ml). De organiske lag ble samlet, tørket (Na2SC«4), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med Heks:EtOAc (4:1) ga sub-tittelforbindelse (5,2 g, 25%). A solution of 3-chloro-5-methoxybenzaldehyde (22.8 g, 134 mmol; see step (i) above) in CH 2 Cl 2 (250 mL) was cooled to 0 °C. Boron tribromide (15.8 mL, 167 mmol) was added dropwise over 15 min. After stirring the reaction mixture for 2 h, H 2 O (50 mL) was added slowly. The solution was then extracted with Et 2 O (2 x 100 mL). The organic layers were combined, dried (Na2SO4), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (4:1) gave the sub-title compound (5.2 g, 25%).

'H NMR (300 MHz, CDC13) 8 9,85 (s, 1H), 7,35 (s,lH), 7,20 (s,lH), 7,10 (s.lH), 3,68 1 H NMR (300 MHz, CDCl 3 ) δ 9.85 (s, 1H), 7.35 (s, 1H), 7.20 (s, 1H), 7.10 (s, 1H), 3.68

(s.lH) (p.lH)

(iii) 3- klor- 5- difluormetoksvbenzaldehvd (iii) 3- chloro- 5-difluoromethoxybenzaldehyde eq

En løsning av 3-klor-5-hydroksybenzaldehyd (7,5 g, 48 mmol; se trinn (ii) ovenfor) i 2-propanol (250 ml) og 30% KOH (100 ml) ble oppvarmet til tilbakeløp. Under omrøring, ble CHCIF2 boblet inn i reaksjonsblandingen i 2 timer. Reaksjonsblandingen ble avkjølt, surgjort med IN HC1 og ekstrahert med EtOAc (2 x 100 ml). De organiske faser ble vasket med saltvann (100 ml), tørket (Na2SC«4), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med Heks:EtOAc (4:1) ga sub-tittelforbindelse (4,6 g, 46%). A solution of 3-chloro-5-hydroxybenzaldehyde (7.5 g, 48 mmol; see step (ii) above) in 2-propanol (250 mL) and 30% KOH (100 mL) was heated to reflux. While stirring, CHCIF2 was bubbled into the reaction mixture for 2 hours. The reaction mixture was cooled, acidified with 1N HCl and extracted with EtOAc (2 x 100 mL). The organic phases were washed with brine (100 mL), dried (Na2SO4), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (4:1) gave the sub-title compound (4.6 g, 46%).

<*>H NMR (300 MHz, CDC13) 8 9,95 (s, 1H), 7,72 (s, 1H), 7,52 (s, 1H), 7,40 (s, 1H), 6,60 (t, 7H-f = 71,1Hz, 1H) <*>H NMR (300 MHz, CDCl 3 ) δ 9.95 (s, 1H), 7.72 (s, 1H), 7.52 (s, 1H), 7.40 (s, 1H), 6, 60 (t, 7H-f = 71.1Hz, 1H)

(iv) Ph( 3- Cl)( 5- OCHF?)-(/ ?. y) CH( OTMS) CN (iv) Ph( 3- Cl)( 5- OCHF?)-(/ ?. y) CH( OTMS) CN

En løsning av 3-klor-5-difluormetoksybenzaldehyd (4,6 g, 22,3 mmol; se trinn (iii) ovenfor) i CH2C12 (200 ml) ble avkjølt til 0°C. Znl2 (1,8 g, 5,6 mmol) og trimetylsilylcyanid (2,8 g, 27,9 mmol) ble tilsatt og reaksjonsblandingen fikk oppvarmes til romtemperatur og omrørt i 15 timer. Blandingen ble delvis konsentrert / vakuum hvilket gir sub-tittelforbindelsen som dets salt, som ble anvendt direkte i trinn (v) nedenfor uten ytterligere rensning eller karakterisering. A solution of 3-chloro-5-difluoromethoxybenzaldehyde (4.6 g, 22.3 mmol; see step (iii) above) in CH 2 Cl 2 (200 mL) was cooled to 0 °C. Zn12 (1.8 g, 5.6 mmol) and trimethylsilyl cyanide (2.8 g, 27.9 mmol) were added and the reaction mixture was allowed to warm to room temperature and stir for 15 hours. The mixture was partially concentrated / vacuum to give the sub-title compound as its salt, which was used directly in step (v) below without further purification or characterization.

(v) Ph( 3- Cl¥5- OCHF7)-(/ ?.^ CH( OH) C( NH) OEt (v) Ph( 3- Cl¥5- OCHF7)-(/ ?.^ CH( OH) C( NH) OEt

Ph(3-Cl)(5-OCHF2)-(/?,S)CH(OTMS)CN (6,82 g, annta 22,3 mmol; se trinn (iv) ovenfor) ble satt dråpevis til HCl/EtOH (500 ml). Reaksjonsblandingen ble omrørt i 15 timer, deretter delvis konsentrert / vakuum hvilket gir sub-tittelforbindelse som dens salt, som ble anvendt i trinn (vi) uten ytterligere rensning eller karakterisering. Ph(3-Cl)(5-OCHF2)-(/?,S)CH(OTMS)CN (6.82 g, assumed 22.3 mmol; see step (iv) above) was added dropwise to HCl/EtOH ( 500 ml). The reaction mixture was stirred for 15 h, then partially concentrated/vacuum to give the sub-title compound as its salt, which was used in step (vi) without further purification or characterization.

(vi) Ph( 3- Cn( 5- OCHFz)-(/ ?. 5) CH( OH) C( Q) OEt (vi) Ph( 3- Cn( 5- OCHFz)-(/ ?. 5) CH( OH) C( Q) OEt

Ph(3-Cl)(5-OCHF2)-(/?,5)CH(OH)C(NH)OEt (6,24 g, anta 22,3 mmol; se trinn (v) ovenfor) ble oppløst i THF (250 ml), 0,5M H2S04Ph(3-Cl)(5-OCHF2)-(/?,5)CH(OH)C(NH)OEt (6.24 g, assume 22.3 mmol; see step (v) above) was dissolved in THF (250 mL), 0.5 M H 2 SO 4

(400 ml) ble tilsatt og reaksjonsblandingen ble omrørt ved 40°C i 65 timer, avkjølt og deretter delvis konsentrert i vakuum for å fjerne mesteparten av THF. Reaksjonsblandingen ble deretter ekstrahert med Et20 (3 x 100 ml), tørket (Na2SC«4), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelse som et fast stoff, som ble anvendt i trinn (vii) uten ytterligere rensning eller karakterisering. (400 mL) was added and the reaction mixture was stirred at 40°C for 65 h, cooled and then partially concentrated in vacuo to remove most of the THF. The reaction mixture was then extracted with Et 2 O (3 x 100 mL), dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give the sub-title compound as a solid, which was used in step (vii) without further purification or characterization.

(vii) Ph( 3- ClX5- OCHF9H/ g. S) CH( OH) C( 0) OH (vii) Ph( 3- ClX5- OCHF9H/ g. S) CH( OH) C( 0) OH

En løsning av Ph(3-Cl)(5-OCHF2)-(/?,5)CH(OH)C(0)OEt (6,25 g, anta 22,3 mmol; se trinn (vi) ovenfor) i 2-propanol (175 ml) og 20% KOH (350 ml) ble omrørt ved romtemperatur 15 timer. Reaksjonen ble deretter delvis konsentrert i vakuum for å fjerne mesteparten av 2-propanol. Den gjenværende blanding ble surgjort med IM H2SC«4, ekstrahert med Et20 (3 x 100 ml), tørket (Na2S04) og konsentrert i vakuum, hvilket gir et fast stoff. Flash kromatografi på silikagel under eluering med CHCl3:MeOH:konsentrert NH4OH (6:3:1) ga ammoniumsaltet av sub-tittelforbindelse. Ammoniumsaltet ble deretter oppløst i en blanding av EtOAc (75 ml) og H20 (75 ml) og surgjort med 2N HC1. Det organiske laget ble separert og vasket med saltvann (50 ml), tørket (Na2SC«4) og konsentrert i vakuum, hvilket gir sub-tittelforbindelsen (3,2 g, 57% fra trinn (iv) til (vii)). A solution of Ph(3-Cl)(5-OCHF2)-(/?,5)CH(OH)C(0)OEt (6.25 g, assume 22.3 mmol; see step (vi) above) in 2-propanol (175 mL) and 20% KOH (350 mL) were stirred at room temperature for 15 h. The reaction was then partially concentrated in vacuo to remove most of the 2-propanol. The remaining mixture was acidified with 1M H 2 SC 4 , extracted with Et 2 O (3 x 100 mL), dried (Na 2 SO 4 ) and concentrated in vacuo to give a solid. Flash chromatography on silica gel eluting with CHCl 3 :MeOH:concentrated NH 4 OH (6:3:1) afforded the ammonium salt of sub-title compound. The ammonium salt was then dissolved in a mixture of EtOAc (75 mL) and H 2 O (75 mL) and acidified with 2N HCl. The organic layer was separated and washed with brine (50 mL), dried (Na 2 SC 4 ) and concentrated in vacuo to give the sub-title compound (3.2 g, 57% from steps (iv) to (vii)).

'H NMR (300 MHz, CD3OD) 8 7,38 (s, 1H), 7,22 (s, 1H), 7,15 (s, 1H), 6,89 (t, 7H-f = 71,1 Hz, 1H), 5,16 (s, 1H) 1 H NMR (300 MHz, CD 3 OD) δ 7.38 (s, 1H), 7.22 (s, 1H), 7.15 (s, 1H), 6.89 (t, 7H-f = 71.1 Hz, 1H), 5.16 (s, 1H)

(viii) Ph( 3- Cn( 5- OCHF?H/ flCH( OH) C( 0) OH ( a) og Ph( 3- Cn( 5- OCHF7)-( S) CH( OAc) C( Q) OH ( b) (viii) Ph( 3- Cn( 5- OCHF?H/ flCH( OH) C( 0) OH ( a) and Ph( 3- Cn( 5- OCHF7)-( S) CH( OAc) C( Q) OH (b)

En blanding av Ph(3-Cl)(5-OCHF2)-(Æ,S)CH(OH)C(0)OH (3,2 g, 12,7 mmol; se trinn (vii) ovenfor) og Lipase PS "Amano" (-2,0 g) i vinylacetat (125 ml) og MTBE (125 ml) ble oppvarmet ved tilbakeløp i 48 timer. Reaksjonsblandingen ble avkjølt, filtrert gjennom Celite® og filterkaken vasket med EtOAc. Filtratet ble konsentrert i vakuum og underkastet flash kromatografi på silikagel under eluering med CHCl3:MeOH:konsentrert NH4OH (6:3:1) hvilket gir ammoniumsalter av sub-tittelforbindelsene (a) og (b). Forbindelse (a) som et salt ble oppløst i H20, surgjort med 2N HC1 og ekstrahert med EtOAc. Det organiske laget ble vasket med saltvann, tørket (Na2SC«4), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelse (a) (1,2 g, 37%). A mixture of Ph(3-Cl)(5-OCHF2)-(Æ,S)CH(OH)C(0)OH (3.2 g, 12.7 mmol; see step (vii) above) and Lipase PS "Amano" (-2.0 g) in vinyl acetate (125 mL) and MTBE (125 mL) was heated at reflux for 48 hours. The reaction mixture was cooled, filtered through Celite® and the filter cake washed with EtOAc. The filtrate was concentrated in vacuo and subjected to flash chromatography on silica gel eluting with CHCl3:MeOH:concentrated NH4OH (6:3:1) to give ammonium salts of the sub-title compounds (a) and (b). Compound (a) as a salt was dissolved in H 2 O, acidified with 2N HCl and extracted with EtOAc. The organic layer was washed with brine, dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give sub-title compound (a) (1.2 g, 37%).

For sub-tittelforbindelse (a) For sub-title connection (a)

'H NMR (300 MHz, CD3OD) 8 7,38 (s, 1H), 7,22 (s, 1H), 7,15 (s, 1H), 6,89 (t, 7H-f = 71,1 Hz, 1H), 5,17 (s, 1H) 1 H NMR (300 MHz, CD 3 OD) δ 7.38 (s, 1H), 7.22 (s, 1H), 7.15 (s, 1H), 6.89 (t, 7H-f = 71.1 Hz, 1H), 5.17 (s, 1H)

(ix) Ph( 3- Cl)( 5- OCHF,)-(/ ?) CH( OH) C( Q)- Aze- Pab( Teoc) (ix) Ph( 3- Cl)( 5- OCHF,)-(/ ?) CH( OH) C( Q)- Aze- Pab( Teoc)

Til en løsning av Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)OH (1,1 g, 4,4 mmol; se trinn (viii) ovenfor) og H-Aze-Pab(Teoc) (se internasjonal patentsøknad WO 00/42059, 2,6 g, 5,7 mmol) i DMF (50 ml) ved 0°C ble tilsatt PyBOP (2,8 g, 5,3 mmol) og collidin (1,3 g, 10,6 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur i ytterligere 15 timer. Reaksjonsblandingen ble konsentrert i vakuum og flash kromatografert på silikagel (3 x), under eluering først med CHCtøEtOH (9:1), deretter med EtOAc:EtOH (20:1) og til slutt under eluering med CH2C12:CH30H (95:5), hvilket gir sub-tittelforbindelsen (1,0 g, 37%) som et hvitt, fast stoff. To a solution of Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)OH (1.1 g, 4.4 mmol; see step (viii) above) and H- Aze-Pab(Teoc) (see International Patent Application WO 00/42059, 2.6 g, 5.7 mmol) in DMF (50 mL) at 0°C was added PyBOP (2.8 g, 5.3 mmol) and collidine (1.3 g, 10.6 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature for an additional 15 hours. The reaction mixture was concentrated in vacuo and flash chromatographed on silica gel (3x), eluting first with CHCl2EtOH (9:1), then with EtOAc:EtOH (20:1) and finally eluting with CH2Cl2:CH3OH (95:5) , giving the sub-title compound (1.0 g, 37%) as a white solid.

'H NMR (300 MHz, CD3OD, blanding av rotamerer) 8 7,79-7,85 (d, J = 8,7 Hz, 2H), 7,15-7,48 (m, 5H), 6,89 og 6,91 (t, 7H-f = 71,1 Hz, 1H), 5,12 og 5,20 (s, 1H), 4,75-4,85 (m, 1H), 3,97-4,55 (m, 6H), 2,10-2,75 (m, 2H), 1,05-1,15 (m, 2H), 0,09 (s, 9H) 1 H NMR (300 MHz, CD 3 OD, mixture of rotamers) δ 7.79-7.85 (d, J = 8.7 Hz, 2H), 7.15-7.48 (m, 5H), 6.89 and 6.91 (t, 7H-f = 71.1 Hz, 1H), 5.12 and 5.20 (s, 1H), 4.75-4.85 (m, 1H), 3.97-4 .55 (m, 6H), 2.10-2.75 (m, 2H), 1.05-1.15 (m, 2H), 0.09 (s, 9H)

MS (m/z)611(M + l)<+>MS (m/z) 611 (M + 1)<+>

(x) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OcBu. Teoc) (x) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OcBu. Teoc)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,051 g, 0,08 mmol; se trinn (ix) ovenfor), ble oppløst i 3 ml acetonitril og 0,062 g Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.051 g, 0.08 mmol; see step (ix) above), was dissolved in 3 ml of acetonitrile and 0.062 g

(0,5 mmol) O-cyklobutylhydroksylamin-hydroklorid ble tilsatt. Blandingen ble oppvarmet ved 70°C i 4,5 timer. Løsningsmidlet ble avdampet og residuet ble fordelt mellom vann og etylacetat. Den vandige fasen ble ekstrahert to ytterligere ganger med etylacetat og den samlede organiske fase ble vasket med vann, saltvann, tørket (Na2SC<4), filtrert og inndampet. Utbytte: 0,054 g (95%). (0.5 mmol) of O-cyclobutylhydroxylamine hydrochloride was added. The mixture was heated at 70°C for 4.5 hours. The solvent was evaporated and the residue was partitioned between water and ethyl acetate. The aqueous phase was extracted twice more with ethyl acetate and the combined organic phase was washed with water, brine, dried (Na2SO4), filtered and evaporated. Yield: 0.054 g (95%).

'H-NMR (400 MHz; CD3OD): 5 8,66-8,50 (m, 1H), 7,45 (d, 2H), 7,29 (m, 3H), 7,15 (m, 2H), 6,88 (t, 1H hoved rotamer), 6,85 (t, 1H mindre rotamer), 5,18 (s,lH hoved rotamer), 5,12 (s, 1H mindre rotamer), 5,16 (m, 1H mindre rotamer), 4,78 (m, 1H hoved rotamer), 4,70 (m, 1H), 4,50-4,30 (m, 3H), 4,19-3,93 (m, 3H), 2,71-2,44 (m, 1H), 2,34-2,11 (m, 5H), 1,78 (m, 1H), 1,62 (m, 1H), 0,96 (m, 2H), 0,01 (s, 9H) 1H-NMR (400 MHz; CD 3 OD): δ 8.66-8.50 (m, 1H), 7.45 (d, 2H), 7.29 (m, 3H), 7.15 (m, 2H ), 6.88 (t, 1H major rotamer), 6.85 (t, 1H minor rotamer), 5.18 (s,lH major rotamer), 5.12 (s, 1H minor rotamer), 5.16 ( m, 1H minor rotamer), 4.78 (m, 1H major rotamer), 4.70 (m, 1H), 4.50-4.30 (m, 3H), 4.19-3.93 (m, 3H), 2.71-2.44 (m, 1H), 2.34-2.11 (m, 5H), 1.78 (m, 1H), 1.62 (m, 1H), 0.96 (m, 2H), 0.01 (s, 9H)

(xi) Ph( 3- Cl¥5- OCHF?)-(/ tfCH( OH) C( 0)- Aze- Pab( OcBu) (xi) Ph( 3- Cl¥5- OCHF?)-(/ tfCH( OH) C( 0)- Aze- Pab( OcBu)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OcBu, Teoc) (0,054 g, Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OcBu, Teoc) (0.054 g,

0,08 mmol; se trinn (x) ovenfor), ble oppløst i 0,5 ml CH2C12 og 0.08 mmol; see step (x) above), was dissolved in 0.5 ml of CH2Cl2 and

3 ml TFA. Reaksjonen fikk forløpe i 60 minutter. TFA ble inndampet og residuet ble renset ved anvendelse av preparativ HPLC. Fraksjonene av interesse ble samlet og frysetørket (2x), hvilket gir 23 mg (54%) av tittelforbindelsen. 3 ml of TFA. The reaction was allowed to proceed for 60 minutes. The TFA was evaporated and the residue was purified using preparative HPLC. The fractions of interest were pooled and freeze-dried (2x), yielding 23 mg (54%) of the title compound.

MS (m/z) 536 (M -1)_; 538 (M + 1)<+>MS (m/z) 536 (M -1)_; 538 (M + 1)<+>

'H-NMR (400 MHz; CD3OD): 8 7,56 (d, 2H), 7,33 (m, 3H), 7,15 (m, 2H), 6,89 (t, 1H hoved rotamer), 6,86 (t, 1H mindre rotamer), 5,18 (s, 1H hoved rotamer; og m, 1H mindre rotamer), 5,11 (s, 1H mindre rotamer), 4,77 (m, 1H hoved rotamer), 4,58 (m, 1H), 4,42 (m, 2H), 4,34 (m, 1H hoved rotamer), 4,15 (m, 1H hoved rotamer), 4,06 (m, 1H mindre rotamer), 3,97 (m, 1H mindre rotamer), 2,66 (m, 1H mindre rotamer), 2,52 (m, 1H hoved rotamer), 2,33-2,25 (m, 3H), 2,01-2,20 (m, 2H), 1,75 (m, 1H), 1,59 (m, 1H) 1H-NMR (400 MHz; CD3OD): δ 7.56 (d, 2H), 7.33 (m, 3H), 7.15 (m, 2H), 6.89 (t, 1H main rotamer), 6.86 (t, 1H minor rotamer), 5.18 (s, 1H major rotamer; and m, 1H minor rotamer), 5.11 (s, 1H minor rotamer), 4.77 (m, 1H major rotamer) , 4.58 (m, 1H), 4.42 (m, 2H), 4.34 (m, 1H major rotamer), 4.15 (m, 1H major rotamer), 4.06 (m, 1H minor rotamer ), 3.97 (m, 1H minor rotamer), 2.66 (m, 1H minor rotamer), 2.52 (m, 1H major rotamer), 2.33-2.25 (m, 3H), 2, 01-2.20 (m, 2H), 1.75 (m, 1H), 1.59 (m, 1H)

<13>C-NMR (100 MHz; CD3OD) (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,4, 172,3,171,9,171,4,152,3 <13>C-NMR (100 MHz; CD3OD) (carbonyl and/or amidine carbon atoms, rotamers) 8 172.4, 172.3,171.9,171.4,152.3

Eksempel 2 Example 2

Ph( 3- Cl¥5- OCHF?)-(/ ftCH( OH) C( CO- Aze- Pab( OH) Ph( 3- Cl¥5- OCHF?)-(/ ftCH( OH) C( CO- Aze- Pab( OH)

(i) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( 0)- Aze- Pab( OH. Teoc) (i) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( 0)- Aze- Pab( OH. Teoc)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,148 g, 0,24 mmol; se Eksempel l(ix) ovenfor), ble oppløst i 9 ml acetonitril og 0,101 g (1,45 mmol) hydroksylamin-hydroklorid ble tilsatt. Blandingen ble oppvarmet ved 70°C i 2,5 timer, filtrert gjennom Celite® og inndampet. Råproduktet (0,145 g; 75% ren) ble anvendt direkte i neste trinn uten ytterligere rensning. Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.148 g, 0.24 mmol; see Example 1(ix) above), was dissolved in 9 ml of acetonitrile and 0.101 g (1.45 mmol) of hydroxylamine hydrochloride was added. The mixture was heated at 70°C for 2.5 hours, filtered through Celite® and evaporated. The crude product (0.145 g; 75% pure) was used directly in the next step without further purification.

(ii) Ph( 3- Cl)( 5- OCHF?)-(/ ?) CH( OH) C( 0)- Aze- Pab( OH) (ii) Ph( 3- Cl)( 5- OCHF?)-(/ ?) CH( OH) C( 0)- Aze- Pab( OH)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OH, Teoc) (0,145 g, 0,23 mmol; se trinn (i) ovenfor), ble oppløst i 0,5 ml CH2C12 og 9 ml TFA. Reaksjonen fikk forløpe i 60 minutter. TFA ble inndampet og residuet ble renset ved anvendelse av preparativ HPLC. Fraksjonene av interesse ble samlet og frysetørket (2x), hvilket gir 72 mg (utbytte over to trinn 62%) av tittelforbindelsen. Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OH, Teoc) (0.145 g, 0.23 mmol; see step (i) above), was dissolved in 0.5 mL CH 2 Cl 2 and 9 mL TFA. The reaction was allowed to proceed for 60 minutes. The TFA was evaporated and the residue was purified using preparative HPLC. The fractions of interest were pooled and freeze-dried (2x), yielding 72 mg (yield over two steps 62%) of the title compound.

MS (m/z) 482 (M - 1)_; 484 (M + 1)<+>MS (m/z) 482 (M - 1)_; 484 (M + 1)<+>

'H-NMR (400 MHz; CD3OD): 8 7,58 (d, 2H), 7,33 (m, 3H), 7,15 (m, 2H), 6,89 (t, 1H hoved rotamer), 6,86 (t, 1H mindre rotamer), 5,18 (s, 1H hoved rotamer; og m, 1H mindre rotamer), 5,12 (s, 1H mindre rotamer), 4,77 (m, 1H hoved rotamer),4,42 (m, 2H), 4,34 (m, 1H hoved rotamer), 4,14 (m, 1H hoved rotamer), 4,06 (m, 1H mindre rotamer), 3,95 (m, 1H mindre rotamer), 2,66 (m, 1H mindre rotamer), 2,50 (m, 1H hoved rotamer), 2,27 (m, 1H hoved rotamer), 2,14 (m, 1H mindre rotamer) 1H-NMR (400 MHz; CD3OD): δ 7.58 (d, 2H), 7.33 (m, 3H), 7.15 (m, 2H), 6.89 (t, 1H main rotamer), 6.86 (t, 1H minor rotamer), 5.18 (s, 1H major rotamer; and m, 1H minor rotamer), 5.12 (s, 1H minor rotamer), 4.77 (m, 1H major rotamer) ,4.42 (m, 2H), 4.34 (m, 1H major rotamer), 4.14 (m, 1H major rotamer), 4.06 (m, 1H minor rotamer), 3.95 (m, 1H minor rotamer), 2.66 (m, 1H minor rotamer), 2.50 (m, 1H major rotamer), 2.27 (m, 1H major rotamer), 2.14 (m, 1H minor rotamer)

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,4,172,3,172,0,171,4 152,3,152,1 <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 172,4,172,3,172,0,171,4 152,3,152,1

Eksempel 3 Example 3

Ph( 3- Cl)( 5- OCHF?)-(/ ?) CH( OH) C( Q)- Aze- Pab Ph( 3- Cl)( 5- OCHF?)-(/ ?) CH( OH) C( Q)- Aze- Pab

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,045 g, 0,074 mmol; se Eksempel l(ix) ovenfor), ble oppløst i 3 ml TFA og fikk reagere i 1 time. TFA ble inndampet og residuet ble fryse tørket fra vann/acetonitril, hvilket gir 0,043 g (100%) av sub-tittelforbindelsen som dens TFA salt. Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.045 g, 0.074 mmol; see Example 1(ix) above), was dissolved in 3 ml of TFA and allowed to react for 1 hour. The TFA was evaporated and the residue was freeze dried from water/acetonitrile to give 0.043 g (100%) of the sub-title compound as its TFA salt.

'H-NMR (400 MHz; CD3OD) rotamerer: 8 7,8-7,75 (m, 2H), 7,55-7,5 (m, 2H), 7,35 (m, 1H, hoved rotamer), 7,31 (m, 1H, mindre rotamer), 7,19 (m, 1H, hoved rotamer), 7,15 (m, 1H), 7,12 (m, 1H, mindre rotamer), 6,89 (t, 1H, hoved rotamer), 6,87 (t, 1H, mindre rotamer), 5,22 (m, 1H, mindre rotamer), 5,20 (s, 1H, hoved rotamer), 5,13 (s, 1H, mindre 1H-NMR (400 MHz; CD3OD) rotamer: δ 7.8-7.75 (m, 2H), 7.55-7.5 (m, 2H), 7.35 (m, 1H, main rotamer) , 7.31 (m, 1H, minor rotamer), 7.19 (m, 1H, major rotamer), 7.15 (m, 1H), 7.12 (m, 1H, minor rotamer), 6.89 ( t, 1H, major rotamer), 6.87 (t, 1H, minor rotamer), 5.22 (m, 1H, minor rotamer), 5.20 (s, 1H, major rotamer), 5.13 (s, 1H, less

rotamer), 4,80 (m, 1H, hoved rotamer), 4,6-4,4 (m, 2H), 4,37 (m, 1H, hoved rotamer), 4,19 (m, 1H, hoved rotamer), 4,07 (m, 1H, mindre rotamer), 3,98 (m, 1H, mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,55 (m, 1H, hoved rotamer), 2,29 (m, 1H, hoved rotamer), 2,15 rotamer), 4.80 (m, 1H, main rotamer), 4.6-4.4 (m, 2H), 4.37 (m, 1H, main rotamer), 4.19 (m, 1H, main rotamer ), 4.07 (m, 1H, minor rotamer), 3.98 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.55 (m, 1H, major rotamer), 2.29 (m, 1H, main rotamer), 2.15

(m, 1H, mindre rotamer) (m, 1H, minor rotamer)

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,6,172,5,172,0,171,7,167,0 <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 172,6,172,5,172,0,171,7,167,0

MS (m/z) 465 (M - 1)_, 467 (M + 1)<+>MS (m/z) 465 (M - 1)_, 467 (M + 1)<+>

Eksempel 4 Example 4

Ph( 3- Cl)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- Pab( COOcPentvl) Ph( 3- Cl)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- Pab( COOcPentvl)

Til en løsning av Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab x TFA To a solution of Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab x TFA

(74 mg, 0,13 mmol; se Eksempel 3 ovenfor) og cyklopentylklorformiat (74 mg, 0.13 mmol; see Example 3 above) and cyclopentyl chloroformate

(44 mg, 0,30 mmol) i metylenklorid (5 ml) ble tilsatt vandig NaOH (0,5 ml, 2M, 1 mmol). Blandingen ble omrørt ved romtemperatur og reaksjonen ble overvåket med HPLC. Etter 2,5 timer, ble vann tilsatt og væskefasene ble separert. Den vandige fasen ble ekstrahert to ganger med metylenklorid. De samlede organiske faser ble tørket (MgS04) og renset på silikagel (først metylenklorid, deretter EtOAc). Etter fjerning av løsningsmidlene / vakuum, det faste residuet ble oppløst i vann/acetonitril og frysetørket, hvilket gir tittelforbindelsen som et hvitt, fast stoff. Utbytte: 33 mg (44%) (44 mg, 0.30 mmol) in methylene chloride (5 mL) was added aqueous NaOH (0.5 mL, 2M, 1 mmol). The mixture was stirred at room temperature and the reaction was monitored by HPLC. After 2.5 hours, water was added and the liquid phases were separated. The aqueous phase was extracted twice with methylene chloride. The combined organic phases were dried (MgSO 4 ) and purified on silica gel (first methylene chloride, then EtOAc). After removal of the solvents / vacuum, the solid residue was dissolved in water/acetonitrile and freeze-dried to give the title compound as a white solid. Yield: 33 mg (44%)

MS (m/z) 579 (M + 1)<+>MS (m/z) 579 (M + 1)<+>

<*>H NMR (400MHz; CD3OD): A 7,79(d, 2H), 7,43-7,30(m, 5H), 7,20-7,ll(m, 2H), 6,90(t, 1H, hoved rotamer), 6,87(t, 1H, mindre rotamer), 5,19(dd, 1H, mindre rotamer), 5,18(s, 1H, hoved rotamer), 5,13(m, 1H), 5,1 l(s, 1H, mindre rotamer), 4,78(dd, 1H, hoved rotamer), 4,45(m, 2H), 4,35(m, 1H, hoved rotamer), 4,16(s, 1H, hoved rotamer), 4,06(s, 1H, mindre rotamer), 3,97(s, 1H, mindre rotamer), 2,68(m, 1H, mindre rotamer), 2,52(s, 1H, hoved rotamer), 2,28(s, 1H, hoved rotamer), 2,16(s, 1H, mindre rotamer), l,90(m, 2H), l,77(m, 4H), l,61(m, 2H) <13>C NMR (karbonyl og/eller amidin protoner; 100 MHz): A 173,6,173,1,172,6,170,3, 165,6 <*>H NMR (400MHz; CD 3 OD): Δ 7.79(d, 2H), 7.43-7.30(m, 5H), 7.20-7.11(m, 2H), 6.90 (t, 1H, major rotamer), 6.87(t, 1H, minor rotamer), 5.19(dd, 1H, minor rotamer), 5.18(s, 1H, major rotamer), 5.13(m , 1H), 5.1 l(s, 1H, minor rotamer), 4.78(dd, 1H, major rotamer), 4.45(m, 2H), 4.35(m, 1H, major rotamer), 4.16(s, 1H, major rotamer), 4.06(s, 1H, minor rotamer), 3.97(s, 1H, minor rotamer), 2.68(m, 1H, minor rotamer), 2, 52(s, 1H, major rotamer), 2.28(s, 1H, major rotamer), 2.16(s, 1H, minor rotamer), 1.90(m, 2H), 1.77(m, 4H ), l.61(m, 2H) <13>C NMR (carbonyl and/or amidine protons; 100 MHz): A 173.6,173.1,172.6,170.3, 165.6

Eksempel 5 Example 5

Ph( 3- Cl)( 5- OCHFz)-(/ ?) CH( OH) C( 0)- Aze- Pab( Z) Ph( 3- Cl)( 5- OCHFz)-(/ ?) CH( OH) C( 0)- Aze- Pab( Z)

Tittelforbindelsen ble fremstilt i henhold til metoden beskrevet i Eksempel 4 ovenfor ved å starte fraPh(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab x TFA (73 mg, 0,13 mmol; se Eksempel 3 ovenfor) og benzylklorformiat (35 mg, 0,21 mmol). Ytterligere rensning ved revers-fase HPLC (0,1M ammoniumacetat/MeCN 40/60) ble nødvendig. De passende fraksjoner ble konsentrert i vakuum og ekstrahert med EtOAc. Utbytte: 24 mg (32%). The title compound was prepared according to the method described in Example 4 above starting from Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab x TFA (73 mg, 0 .13 mmol; see Example 3 above) and benzyl chloroformate (35 mg, 0.21 mmol). Further purification by reverse-phase HPLC (0.1M ammonium acetate/MeCN 40/60) was required. The appropriate fractions were concentrated in vacuo and extracted with EtOAc. Yield: 24 mg (32%).

MS (m/z) 602 (M + 1)<+>MS (m/z) 602 (M + 1)<+>

<!>H NMR (400MHz; CD3OD): A 7,80(d, 2H), 7,43-7,25(m, 8H), 7,20-7,10(m, 2H), 6,90(t, 1H, hoved rotamer), 6,88(t, 1H, mindre rotamer), 5,18(dd, 1H, mindre rotamer), 5,18(s, 2H), 5,17(s, 1H, rotamer), 5,1 l(s, 1H, rotamer), 4,78(dd, 1H, hoved rotamer), 4,45(m, 2H), 4,34(m, 1H, hoved rotamer), 4,15(s, 1H, hoved rotamer), 4,06(s, 1H, mindre rotamer), 3,97(s, 1H, mindre rotamer), 2,66(m, 1H, mindre rotamer), 2,5l(s, 1H, hoved rotamer), 2,27(s, 1H, hoved rotamer), 2,15(s, 1H, mindre rotamer) <!>H NMR (400MHz; CD3OD): Δ 7.80(d, 2H), 7.43-7.25(m, 8H), 7.20-7.10(m, 2H), 6.90 (t, 1H, major rotamer), 6.88(t, 1H, minor rotamer), 5.18(dd, 1H, minor rotamer), 5.18(s, 2H), 5.17(s, 1H, rotamer), 5.1 l(s, 1H, rotamer), 4.78(dd, 1H, main rotamer), 4.45(m, 2H), 4.34(m, 1H, main rotamer), 4, 15(s, 1H, major rotamer), 4.06(s, 1H, minor rotamer), 3.97(s, 1H, minor rotamer), 2.66(m, 1H, minor rotamer), 2.5l( s, 1H, major rotamer), 2.27(s, 1H, major rotamer), 2.15(s, 1H, minor rotamer)

<13>C NMR (karbonyl og/eller amidin protoner; 100MHz): A 173,6,173,1,172,6,170,5, 164,9 <13>C NMR (carbonyl and/or amidine protons; 100MHz): A 173.6,173.1,172.6,170.5, 164.9

Eksempel 6 Example 6

Ph( 3- Cl¥5- OCFO-(/ g) CH( OH) C( Q)- Aze- Pab x TFA Ph( 3- Cl¥5- OCFO-(/ g) CH( OH) C( Q)- Aze- Pab x TFA

(i) 2- nitro- 5- trifluormetoksvbenzosvre (i) 2-nitro-5-trifluoromethoxybenzoic acid

Til en løsning av 3-trifluormetoksybenzosyre (49,0 g, 0,24 mol) i svovelsyre (500 ml) at mindre enn 0°C (is-MeOH bad) ble tilsatt en løsning av kaliumnitrat (31,3 g, 0,31 mol) i svovelsyre (200 ml) over 20 minutter. Den resulterende løsning ble omrørt ved 0°C i 2 timer, deretter oppvarmet til romtemperatur og omrørt i 18 timer. Reaksjonsblandingen ble hellet i is og den resulterende sure løsning ble ekstrahert med EtOAc (5x). De samlede organiske lag ble vasket med H20 (lx), saltvann (2x), H20 (lx) og saltvann (lx), tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (65,7 g) som et fast stoff forurenset med HO Ac. Rå sub-tittelforbindelse ble oppløst i EtOAc og toluen og konsentrert i vakuum, hvilket gir et HOAc fritt fast stoff (58,4 g, 97%) som ble anvendt i neste trinn uten ytterligere rensning. To a solution of 3-trifluoromethoxybenzoic acid (49.0 g, 0.24 mol) in sulfuric acid (500 mL) at less than 0°C (ice-MeOH bath) was added a solution of potassium nitrate (31.3 g, 0, 31 mol) in sulfuric acid (200 ml) over 20 minutes. The resulting solution was stirred at 0°C for 2 hours, then warmed to room temperature and stirred for 18 hours. The reaction mixture was poured into ice and the resulting acidic solution was extracted with EtOAc (5x). The combined organic layers were washed with H 2 O (lx), brine (2x), H 2 O (lx) and brine (lx), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound (65.7 g) as a solid contaminated with HO Ac. Crude sub-title compound was dissolved in EtOAc and toluene and concentrated in vacuo to give an HOAc free solid (58.4 g, 97%) which was used in the next step without further purification.

<*>H NMR (300 MHz, CDC13): A 10,10 (br s, 1H), 8,02 (d, 1H, / = 8 Hz), 7,69 (d, 1H, J = 2 Hz), 7,54 (dd, 1H, J = 2 Hz, J = 8 Hz) <*>H NMR (300 MHz, CDCl 3 ): Δ 10.10 (br s, 1H), 8.02 (d, 1H, / = 8 Hz), 7.69 (d, 1H, J = 2 Hz) , 7.54 (dd, 1H, J = 2 Hz, J = 8 Hz)

(ii) 2- amino- 5- trifluormetoksvbenzosvre (ii) 2-amino-5-trifluoromethoxybenzoic acid

Til en løsning av 2-nitro-5-trifluormetoksybenzosyre (56,8 g, 0,23 mol; se trinn (i) ovenfor) i EtOH (1000 ml) ble tilsatt 10% Pd/C (5,7 g). Den resulterende løsning ble spylt med H2 i 5 timer, filtrert gjennom Celite® og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (49,7 g, 98%) som et fast stoff som ble anvendt i neste trinn uten ytterligere rensning. To a solution of 2-nitro-5-trifluoromethoxybenzoic acid (56.8 g, 0.23 mol; see step (i) above) in EtOH (1000 mL) was added 10% Pd/C (5.7 g). The resulting solution was flushed with H 2 for 5 h, filtered through Celite® and concentrated in vacuo to give crude sub-title compound (49.7 g, 98%) as a solid which was used in the next step without further purification.

<*>H NMR (300 MHz, CD3OD): A 7,66 (m, 1H), 7,17 (d, 1H, J = 8 Hz), 6,77 (d, 1H, J = 8 Hz) <*>H NMR (300 MHz, CD3OD): A 7.66 (m, 1H), 7.17 (d, 1H, J = 8 Hz), 6.77 (d, 1H, J = 8 Hz)

(iii) 2- amino- 3- klor- 5- trifluormetoksvbenzosvre (iii) 2-amino-3-chloro-5-trifluoromethoxybenzoic acid

Til en løsning av 2-amino-5-trifluormetoksybenzosyre (49,0 g, 0,22 mol; se trinn (ii) ovenfor) i HOAc (1200 ml) ble langsomt tilsatt sulfurylklorid (41,8 g, 0,31 mol). Gassutvikling ble observert. Den resulterende heterogene blanding ble omrørt ved romtemperatur i 1 time. Ytterligere HOAc (300 ml) ble satt til for å lette omrøringen, fulgt av sulfurylklorid i 5 ml porsjoner inntil utgangsmaterialet ble oppbrukt basert på TLC analyse. Reaksjonen ble konsentrert i vakuum, hvilket gir faste stoffer som ble spylt på en rotasjonsinndamper med EtOAc (2x) fulgt av Et20 (lx) for å fjerne HOAc. De resulterende faste stoffer ble videre tørket, hvilket gir HC1 salt av rå sub-tittelforbindelse (60,5 g, 94%), som ble anvendt i neste trinn uten ytterligere rensning. To a solution of 2-amino-5-trifluoromethoxybenzoic acid (49.0 g, 0.22 mol; see step (ii) above) in HOAc (1200 mL) was slowly added sulfuryl chloride (41.8 g, 0.31 mol) . Gas evolution was observed. The resulting heterogeneous mixture was stirred at room temperature for 1 hour. Additional HOAc (300 mL) was added to facilitate stirring, followed by sulfuryl chloride in 5 mL portions until the starting material was consumed based on TLC analysis. The reaction was concentrated in vacuo to give solids which were flushed on a rotary evaporator with EtOAc (2x) followed by Et 2 O (1x) to remove HOAc. The resulting solids were further dried to give the HCl salt of crude sub-title compound (60.5 g, 94%), which was used in the next step without further purification.

'H NMR (300 MHz, CD3OD): A 7,72 (s, 1H), 7,44 (s, 1H), 7,22 (s, exchangeables) 1 H NMR (300 MHz, CD 3 OD): Δ 7.72 (s, 1H), 7.44 (s, 1H), 7.22 (s, exchangeables)

(iv) 3- klor- 5- trifluormetoksvbenzosvre (iv) 3-chloro-5-trifluoromethoxybenzoic acid

Til en løsning av 2-amino-3-klor-5-trifluormetoksybenzosyre (60,5 g, anta 0,22 mol; se trinn (iii) ovenfor) i 1,4-dioksan (1000 ml) ble tilsatt 6N HC1 (750 ml). Noen organiske stoffer gikk ut av løsningen som olje. Dioksanløsningen ble avkjølt til mindre enn 0°C (is-MeOH bad). En løsning av natriumnitritt (18,2 g, 0,26 mol) i H20 (250 ml) ble tilsatt over 15 minutter via en tilsetningstrakt. Den resulterende løsning ble omrørt i 45 min. Hypofosforsyrling (221,5 ml 50 vekt% i H20, 291,2 g, To a solution of 2-amino-3-chloro-5-trifluoromethoxybenzoic acid (60.5 g, assume 0.22 mol; see step (iii) above) in 1,4-dioxane (1000 mL) was added 6N HCl (750 ml). Some organic substances came out of the solution as oil. The dioxane solution was cooled to less than 0°C (ice-MeOH bath). A solution of sodium nitrite (18.2 g, 0.26 mol) in H 2 O (250 mL) was added over 15 min via an addition funnel. The resulting solution was stirred for 45 min. Hypophosphorous acid (221.5 ml 50% by weight in H20, 291.2 g,

2,20 mol) ble tilsatt langsomt via en tilsetningstrakt. Løsningen ble omrørt ved 0°C i 1,5 timer, deretter oppvarmet til romtemperatur (gassutvikling ble observert) og omrørt i 18 timer. Rå løsning ble overført til en separasjonstrakt og ekstrahert med Et20 (4x). De samlede organiske lag ble ekstrahert med vandig NaHCC<3 (3x). Det basiske vandige lag ble forsiktig surgjort med 6N HC1 og ekstrahert med CH2C12 (3x). CH2C12 ekstrakter ble tørket (Na2SC«4), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (26,5 g, 46% fra 3-trifluormetoksybenzosyre) som et fast stoff som ble anvendt i neste trinn uten ytterligere rensning. 2.20 mol) was added slowly via an addition funnel. The solution was stirred at 0°C for 1.5 hours, then warmed to room temperature (gas evolution was observed) and stirred for 18 hours. Crude solution was transferred to a separatory funnel and extracted with Et 2 O (4x). The combined organic layers were extracted with aqueous NaHCC<3 (3x). The basic aqueous layer was gently acidified with 6N HCl and extracted with CH 2 Cl 2 (3x). CH 2 Cl 2 extracts were dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give crude sub-title compound (26.5 g, 46% from 3-trifluoromethoxybenzoic acid) as a solid which was used in the next step without further purification.

'H NMR (300 MHz, CD3OD): A 7,98 (s, 1H), 7,83 (s, 1H), 7,58 (s, 1H) 1 H NMR (300 MHz, CD 3 OD): Δ 7.98 (s, 1H), 7.83 (s, 1H), 7.58 (s, 1H)

(v) 3- klor- 5- trifluormetoksvbenzvlalkohol (v) 3-chloro-5-trifluoromethoxybenzoyl alcohol

Til en løsning av 3-klor-5-trifluormetoksybenzosyre (22,5 g, To a solution of 3-chloro-5-trifluoromethoxybenzoic acid (22.5 g,

93,5 mmol; se trinn (iv) ovenfor) i vannfri THF (1200 ml) under en N2 atmosfære ved romtemperatur ble tilsatt en løsning av BH3»THF kompleks (140 ml IM i THF; 140,3 mmol). Løsningen ble tilbakeløpskokt i 2 timer, avkjølt til romtemperatur og omrørt i 18 timer, quenched forsiktig med H20 og konsentrert 1 vakuum for å fjerne mesteparten av THF. Residuet ble fortynnet med EtOAc og de organiske faser ble vasket med saltvann (3x), tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (21,2 g, 100%) som en olje som ble anvendt uten ytterligere rensning. 93.5 mmol; see step (iv) above) in anhydrous THF (1200 mL) under a N 2 atmosphere at room temperature was added a solution of BH 3 »THF complex (140 mL IM in THF; 140.3 mmol). The solution was refluxed for 2 h, cooled to room temperature and stirred for 18 h, carefully quenched with H 2 O and concentrated 1 vacuum to remove most of the THF. The residue was diluted with EtOAc and the organic phases were washed with brine (3x), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound (21.2 g, 100%) as an oil which was used without further purification.

<!>H NMR (300 MHz, CDCI3): A 7,33 (s, 1H), 7,17 (s, 1H), 7,14 (s, 1H), 4,72 (s, 2H), 2,05 <!>H NMR (300 MHz, CDCl 3 ): Δ 7.33 (s, 1H), 7.17 (s, 1H), 7.14 (s, 1H), 4.72 (s, 2H), 2 ,05

(br s, 1H) (br s, 1H)

(vi) 3- klor- 5- trifluormetoksvbenzaldehvd (vi) 3- chloro- 5- trifluoromethoxybenzaldehyde eq

En løsning av DMSO (16,1 g, 205,9 mmol) i vannfri CH2C12 (300 ml) ble avkjølt til -78°C. Oksalylklorid (13,1 g, 103,0 mmol) ble tilsatt langsomt via en sprøyte (gassutvikling ble observert). Den resulterende løsning ble omrørt ved -78°C i 15 minutter. En løsning av 3-klor-5-tirfluormetoksybenzylalkohol (21,2 g, 93,6 mmol; se trinn (v) ovenfor) i CH2C12 (200 ml) ble tilsatt via en tilsetningstrakt over en periode på 15 minutter. Den uklare løsningen ble omrørt ved -78°C i 40 minutter og DIPEA (60,5 g, 468,0 mmol) ble tilsatt via en tilsetningstrakt over 10 minutter. Den resulterende homogene løsning ble omrørt ved -78°C i 1,5 timer, deretter oppvarmet til romtemperatur og omrørt 18 timer. Rå løsning ble konsentrert i vakuum, residuet fortynnet med EtOAc og vasket med H2O (lx), 2N HC1 (lx), saltvann (lx), vandig NaHC03 (lx) og saltvann (lx). De organiske faser ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (19,9 g, 95%) som ble anvendt i neste trinn uten ytterligere rensning. A solution of DMSO (16.1 g, 205.9 mmol) in anhydrous CH 2 Cl 2 (300 mL) was cooled to -78 °C. Oxalyl chloride (13.1 g, 103.0 mmol) was added slowly via syringe (gas evolution was observed). The resulting solution was stirred at -78°C for 15 minutes. A solution of 3-chloro-5-thyrfluoromethoxybenzyl alcohol (21.2 g, 93.6 mmol; see step (v) above) in CH 2 Cl 2 (200 mL) was added via an addition funnel over a period of 15 minutes. The cloudy solution was stirred at -78°C for 40 min and DIPEA (60.5 g, 468.0 mmol) was added via an addition funnel over 10 min. The resulting homogeneous solution was stirred at -78°C for 1.5 hours, then warmed to room temperature and stirred for 18 hours. Crude solution was concentrated in vacuo, the residue diluted with EtOAc and washed with H 2 O (lx), 2N HCl (lx), brine (lx), aqueous NaHCO 3 (lx) and brine (lx). The organic phases were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound (19.9 g, 95%) which was used in the next step without further purification.

<*>H NMR (300 MHz, CDCI3): A 10,00 (s, 1H), 7,83 (s, 1H), 7,66 (s, 1H), 7,51 (s, 1H) <*>H NMR (300 MHz, CDCl 3 ): Δ 10.00 (s, 1H), 7.83 (s, 1H), 7.66 (s, 1H), 7.51 (s, 1H)

(vii) Ph( 3- Cl)( 5- OCFj)-(/ ?. 5) CH( OTMS) CN (vii) Ph( 3- Cl)( 5- OCFj)-(/ ?. 5) CH( OTMS) CN

Til en løsning av 3-klor-5-trifluormetoksybenzaldehyd (19,9 g, To a solution of 3-chloro-5-trifluoromethoxybenzaldehyde (19.9 g,

88,6 mmol; se trinn (vi) ovenfor) i CH2C12 (600 ml) ved 0°C ble tilsatt Znl2 (1,4 g, 4,4 mmol) og trimetylsilylcyanid (9,7 g, 97,5 mmol). Etter omrøring ved 0°C i 1,5 timer og ved romtemperatur i 2 timer, viste TLC analyse bare utgangsmaterialet. Znl2 ble tilsatt porsjonsvis inntil reaksjonen forløp (over 30,0 g Znl2 ble tilsatt totalt). Etter omrøring ved romtemperatur i 18 timer, ble reaksjonen stanset med vann og de organiske faser ble separert. De organiske faser ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (27,7 g, 96%) som en væske som ble anvendt uten ytterligere rensning. 88.6 mmol; see step (vi) above) in CH 2 Cl 2 (600 mL) at 0 °C was added Zn 2 (1.4 g, 4.4 mmol) and trimethylsilyl cyanide (9.7 g, 97.5 mmol). After stirring at 0°C for 1.5 hours and at room temperature for 2 hours, TLC analysis showed only the starting material. Znl2 was added in portions until the reaction proceeded (more than 30.0 g of Znl2 was added in total). After stirring at room temperature for 18 hours, the reaction was quenched with water and the organic phases were separated. The organic phases were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound (27.7 g, 96%) as a liquid which was used without further purification.

<J>H NMR (300 MHz, CDC13): A 7,43 (s, 1H), 7,28 (s, 1H), 7,25 (s, 1H), 5,49 (s, 1H), 0,38 <J>H NMR (300 MHz, CDCl 3 ): Δ 7.43 (s, 1H), 7.28 (s, 1H), 7.25 (s, 1H), 5.49 (s, 1H), 0 ,38

(s, 9H) (pp, 9H)

(viii) Ph( 3- Cl)( 5- OCF^-(/ g. 5) CH( OH) C( Q) OH (viii) Ph( 3- Cl)( 5- OCF^-(/ g. 5) CH( OH) C( Q) OH

En suspensjon av Ph(3-Cl)(5-OCF3)-(Æ,S)CH(OTMS)CN (27,7 g, A suspension of Ph(3-Cl)(5-OCF3)-(Æ,S)CH(OTMS)CN (27.7 g,

85,6 mmol; se trinn (vii) ovenfor) i konsentrert HC1 (300 ml) ble tilbakeløpskokt i 3 timer. Den resulterende brune heterogene blanding ble avkjølt til romtemperatur og ekstrahert med Et20 (2x). De innledende organiske lag ble ekstrahert med 2N NaOH (2x), deretter ble basiske lag surgjort med 2N HC1 og ekstrahert med EtiO. Et20 ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (4,9 g, 21%). TLC analyse av de innledende organiske lag viste at sub-tittelforbindelsen fortsatt var til stede slik at basisk ekstraksjon/surgjøring ble gjentatt ved anvendelse av 6N NaOH, hvilket gir 85.6 mmol; see step (vii) above) in concentrated HCl (300 mL) was refluxed for 3 h. The resulting brown heterogeneous mixture was cooled to room temperature and extracted with Et 2 O (2x). The initial organic layers were extracted with 2N NaOH (2x), then basic layers were acidified with 2N HCl and extracted with EtiO. Et 2 O was dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound (4.9 g, 21%). TLC analysis of the initial organic layers showed that the sub-title compound was still present so basic extraction/acidification was repeated using 6N NaOH, giving

ytterligere rå sub-tittelforbindelse (2,8 g, 12%). TLC analyse av de innledende organiske lag viste at sub-tittelforbindelsen fortsatt var til stede slik at de organiske faser ble tørket (Na2S04) og konsentrert i vakuum, hvilket gir natriumsaltet av sub-tittelforbindelse (18,3 g) som en olje. Saltet ble deretter gjenoppløst i Et20 og de organiske faser surgjort med 2N HC1 og vasket med saltvann. Det resulterende organiske lag ble tørket (Na2S04), additional crude sub-title compound (2.8 g, 12%). TLC analysis of the initial organic layers showed that the sub-title compound was still present so the organic phases were dried (Na 2 SO 4 ) and concentrated in vacuo to give the sodium salt of the sub-title compound (18.3 g) as an oil. The salt was then redissolved in Et2O and the organic phases acidified with 2N HCl and washed with brine. The resulting organic layer was dried (Na 2 SO 4 ),

behandlet med aktivert trekull, filtrert gjennom Celite® og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (14,3 g, 62%) som et fast stoff som ble anvendt i neste trinn uten ytterligere rensning. treated with activated charcoal, filtered through Celite® and concentrated in vacuo to give crude sub-title compound (14.3 g, 62%) as a solid which was used in the next step without further purification.

<*>H NMR (300 MHz, CD3OD): A 7,53 (s, 1H), 7,38 (s, 1H), 7,29 (s, 1H), 5,23 (s, 1H) <*>H NMR (300 MHz, CD3OD): Δ 7.53 (s, 1H), 7.38 (s, 1H), 7.29 (s, 1H), 5.23 (s, 1H)

(ix) Ph( 3- Cl)( 5- OCFO-(/ g) CH( OH) C( C0OH ( a) oe Ph( 3- Cl)( 5- OCFO-( S) CH( OAc) C( Q) OH ( b) (ix) Ph( 3- Cl)( 5- OCFO-(/ g) CH( OH) C( C0OH ( a) oe Ph( 3- Cl)( 5- OCFO-( S) CH( OAc) C( Q ) OH (b)

En blanding av Ph(3-Cl)(5-OCF3)-(Æ,S)CH(OH)C(0)OH (7,7 g, 28,5 mmol; se trinn (viii) ovenfor) og Lipase PS "Amano" (3,8 g) i MTBE (100 ml) og vinylacetat (50 ml) ble omrørt ved 60°C i 26 timer. Reaksjonsblandingen ble avkjølt og filtrert gjennom Celite® og filterkaken vasket med EtOAc. De samlede organiske lag ble konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:MeOH:konsentrert NH4OH (6:3:1) ga en blanding av ammoniumsalter av sub-tittelforbindelse (a) og sub-tittelforbindelse (b) (6,7 g) og en ren prøve av ammoniumsaltet av sub-tittelforbindelse (a) A mixture of Ph(3-Cl)(5-OCF3)-(Æ,S)CH(OH)C(0)OH (7.7 g, 28.5 mmol; see step (viii) above) and Lipase PS "Amano" (3.8 g) in MTBE (100 mL) and vinyl acetate (50 mL) was stirred at 60°C for 26 h. The reaction mixture was cooled and filtered through Celite® and the filter cake washed with EtOAc. The combined organic layers were concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl3:MeOH:conc NH4OH (6:3:1) gave a mixture of ammonium salts of sub-title compound (a) and sub-title compound (b) (6.7 g) and a pure sample of the ammonium salt of sub-title compound (a)

(1,2 g) med mindre enn 95% e.e. De respektive fraksjoner ble oppløst i Et20 og vasket med 2N HC1 (lx) og saltvann (lx), tørket (Na2S04), filtrert og konsentrert, hvilket gir de tilsvarende karboksylsyrer (6,7 g og 1,1 g henholdsvis). Disse fraksjoner ble deretter separat igjen underkastet spaltning og igjen renset som nødvendig via kromatografi på silikagel under eluering med CHCl3:MeOH:konsentrert NH4OH (6:3:1 eller 75:20:5 eller 145:45:10) etter behov. Renset sub-tittelforbindelse (a) ble surgjort med vandig HC1 eller vandig sitronsyre før ytterligere anvendelse. Ammoniumsaltet av sub-tittelforbindelse (b) ble anvendt uten karakterisering. (1.2 g) with less than 95% e.e. The respective fractions were dissolved in Et 2 O and washed with 2N HCl (lx) and brine (lx), dried (Na 2 SO 4 ), filtered and concentrated to give the corresponding carboxylic acids (6.7 g and 1.1 g respectively). These fractions were then separately again subjected to digestion and again purified as necessary via chromatography on silica gel eluting with CHCl3:MeOH:concentrated NH4OH (6:3:1 or 75:20:5 or 145:45:10) as required. Purified sub-title compound (a) was acidified with aqueous HCl or aqueous citric acid before further use. The ammonium salt of sub-title compound (b) was used without characterization.

For sub-tittelforbindelse (a) For sub-title connection (a)

<*>H NMR (300 MHz, CD3OD): A 7,53 (s, 1H), 7,38 (s, 1H), 7,29 (s, 1H), 5,23 (s, 1H) <*>H NMR (300 MHz, CD3OD): Δ 7.53 (s, 1H), 7.38 (s, 1H), 7.29 (s, 1H), 5.23 (s, 1H)

,<3>C NMR (75 MHz, CD3OD): A 174,9,150,9,145,4,136,3,126,8,122,0,120,6,118,9, 72,9 ,<3>C NMR (75 MHz, CD3OD): A 174.9,150,9,145,4,136,3,126,8,122,0,120,6,118,9, 72.9

MS (m/z) 269 (M -1)" MS (m/z) 269 (M -1)"

(x) Ph( 3- CI)( 5- OCFO-(/ g) CH( OH) C( 0)- Aze- Pab( Teoc) (x) Ph( 3- CI)( 5- OCFO-(/ g) CH( OH) C( 0)- Aze- Pab( Teoc)

En løsning av Ph(3-Cl)(5-OCF3)-(Æ)CH(OH)C(0)OH (0,73 g, 2,70 mmol; se trinn (ix) ovenfor) i DMF (40 ml) under en nitrogen-atmosfære ble avkjølt til 0°C. Til løsningen ble satt H-Aze-Pab(Teoc) (1,46 g, A solution of Ph(3-Cl)(5-OCF3)-(Æ)CH(OH)C(0)OH (0.73 g, 2.70 mmol; see step (ix) above) in DMF (40 mL ) under a nitrogen atmosphere was cooled to 0°C. To the solution was added H-Aze-Pab(Teoc) (1.46 g,

3,24 mmol), collidin (0,82 g, 6,75 mmol) og PyBOP (1,83 g, 3,51 mmol). Løsningen ble omrørt ved 0°C i 2 timer, oppvarmet til romtemperatur og omrørt i 18 timer, behandlet med vann og konsentrert / vakuum. Residuet ble fortynnet med EtOAc og vasket med H2O (lx), vandig NaHC03 (lx), vandig sitronsyre (lx) og saltvann (lx), tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse. Flash kromatografi på silikagel (2x) under eluering med EtOAcrMeOH (30:1) deretter CH2Cl2:MeOH (93:7) ga sub-tittelforbindelsen (0,73 g, 43%) som et knusbart skum. 3.24 mmol), collidine (0.82 g, 6.75 mmol) and PyBOP (1.83 g, 3.51 mmol). The solution was stirred at 0°C for 2 hours, warmed to room temperature and stirred for 18 hours, treated with water and concentrated/vacuum. The residue was diluted with EtOAc and washed with H 2 O (lx), aqueous NaHCO 3 (lx), aqueous citric acid (lx) and brine (lx), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound. Flash chromatography on silica gel (2x) eluting with EtOAcrMeOH (30:1) then CH 2 Cl 2 :MeOH (93:7) gave the sub-title compound (0.73 g, 43%) as a crushable foam.

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer): A 7,78-7,82 (d, 2H, J = 8 Hz), 7,25-7,54 (m, 5H), 5,25 og 5,16 (s, 1H), 5,22 og 4,79 (m, 1H), 3,92-4,58 (m, 6H), 2,20-2,76 (m, 2H), 1,04-1,13 (m, 2H), 0,08 (s, 9H) <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers): A 7.78-7.82 (d, 2H, J = 8 Hz), 7.25-7.54 (m, 5H), 5 .25 and 5.16 (s, 1H), 5.22 and 4.79 (m, 1H), 3.92-4.58 (m, 6H), 2.20-2.76 (m, 2H) , 1.04-1.13 (m, 2H), 0.08 (s, 9H)

MS (m/z) 629 (M + 1)<+>MS (m/z) 629 (M + 1)<+>

(xi) Ph( 3- Cl)( 5- OCFO-(/ g) CH( OH) C( Q)- Aze- Pab (xi) Ph( 3- Cl)( 5- OCFO-(/ g) CH( OH) C( Q)- Aze- Pab

Trifluoreddiksyre (1,0 ml) ble satt til en omrørt is/vann-avkjølt løsning av Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (101 mg; 160 umol; se trinn (x) ovenfor), i metylenklorid (10 ml). Kjølebadet ble fjernet etter 1 time. Etter 1,5 timer ved romtemperatur, ble acetonitril (30 ml) tilsatt og løsningsmidlene ble forsiktig fjernet under redusert trykk. Residuet ble oppløst i vann og fryse tørket, hvilket gir 90 mg (92%) av tittelforbindelsen som dens TFA salt. Trifluoroacetic acid (1.0 mL) was added to a stirred ice/water-cooled solution of Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (101 mg; 160 µmol; see step (x) above), in methylene chloride (10 mL). The cooling bath was removed after 1 hour. After 1.5 h at room temperature, acetonitrile (30 mL) was added and the solvents were carefully removed under reduced pressure. The residue was dissolved in water and freeze dried, yielding 90 mg (92%) of the title compound as its TFA salt.

MS (m/z) 483 (M -1)"; 485 (M + 1)<+>MS (m/z) 483 (M -1)"; 485 (M + 1)<+>

'H NMR (300 MHz; CD3OD): (kompleks på grunn av diastereomerene/rotamerer): A 7,70-7,80 (m, 2H), 7,45-7,58 (m, 3H), 7,24-7,38 (m, 2H), 5,26 (s, 1H), 5,17 (m, 1H, mindre rotamer), 4,82 (m, 1H, hoved rotamer), 4,35-4,6 (m, 3H), 4,22 (m, 1H, hoved rotamer), 1 H NMR (300 MHz; CD 3 OD): (complex due to the diastereomers/rotamers): A 7.70-7.80 (m, 2H), 7.45-7.58 (m, 3H), 7.24 -7.38 (m, 2H), 5.26 (s, 1H), 5.17 (m, 1H, minor rotamer), 4.82 (m, 1H, major rotamer), 4.35-4.6 (m, 3H), 4.22 (m, 1H, main rotamer),

3,92-4,12 (m, 2H, mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,55 (m, 1H, hoved rotamer), 2,30 (m, 1H, hoved rotamer), 2,16 (m, 1H, mindre rotamer) 3.92-4.12 (m, 2H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.55 (m, 1H, major rotamer), 2.30 (m, 1H, major rotamer ), 2.16 (m, 1H, minor rotamer)

<13>C NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer): A 173,7,173,4,173,0,172,8, 168,1 <13>C NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers): A 173.7,173.4,173.0,172.8, 168.1

Eksempel 7 Example 7

Ph( 3- Cl)( 5- OCFO-(/ ?) CH( OH) C( Q)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCFO-(/ ?) CH( OH) C( Q)- Aze- Pab( OMe)

HATU (71 mg; 0,19 mmol) ble satt til en omrørt is/vann-avkjølt løsning av Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)OH (39 mg; 0,14 mmol; se Eksempel 6(ix) ovenfor) i DMF (3 ml). Etter 30 minutter, ble en løsning av H-Aze-Pab(OMe) x 2HC1 (69 mg; 0,21 mmol; se internasjonal patentsøknad WO 00/42059) og 2,4,6-collidin (0,080 ml; 0,58 mmol) i DMF (1,5 ml) tilsatt. Reaksjonsblandingen ble latt stå natten over og temperaturen fikk øke langsomt til omgivelsestemperatur. Løsningsmidlene ble fjernet i vakuum og råproduktet ble renset ved anvendelse av revers-fase HPLC (acetonitril: 0,1M vandig ammoniumacetat) hvilket gir, etter frysetørking de passende fraksjoner, tittelforbindelsen (61 mg, 97%) som et fargeløst, fast stoff. HATU (71 mg; 0.19 mmol) was added to a stirred ice/water-cooled solution of Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)OH (39 mg ; 0.14 mmol; see Example 6(ix) above) in DMF (3 mL). After 30 min, a solution of H-Aze-Pab(OMe) x 2HCl (69 mg; 0.21 mmol; see International Patent Application WO 00/42059) and 2,4,6-collidine (0.080 mL; 0.58 mmol) in DMF (1.5 mL) added. The reaction mixture was allowed to stand overnight and the temperature allowed to rise slowly to ambient temperature. The solvents were removed in vacuo and the crude product was purified using reverse-phase HPLC (acetonitrile: 0.1M aqueous ammonium acetate) to give, after freeze-drying the appropriate fractions, the title compound (61 mg, 97%) as a colorless solid.

MS (m/z) 513 (M -1)_, 515 (M + 1)<+>MS (m/z) 513 (M -1)_, 515 (M + 1)<+>

'H NMR (500 MHz; CD3OD): A 7,97 (bt, 1H), 7,53 (d, 2H), 7,27 (t, 1H), 7,22 (d, 2H), 7,19 (t, 1H), 7,11 (t, 2H), 6,77 (s, 1H), 4,92 (s, 1H), 4,9 (bs, 3H), 4,81 (m, 2H), 4,40 (m, 2H), 4,09 (m, 1H) 3,87 (s, 3H), 2,58 (m, 1H), 2,37(m, 1H) 1 H NMR (500 MHz; CD 3 OD): Δ 7.97 (bt, 1H), 7.53 (d, 2H), 7.27 (t, 1H), 7.22 (d, 2H), 7.19 (t, 1H), 7.11 (t, 2H), 6.77 (s, 1H), 4.92 (s, 1H), 4.9 (bs, 3H), 4.81 (m, 2H) , 4.40 (m, 2H), 4.09 (m, 1H) 3.87 (s, 3H), 2.58 (m, 1H), 2.37(m, 1H)

<13>C NMR (125 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer): A 171,8,169,9, 156,8 <13>C NMR (125 MHz; CD3OD): (carbonyl and/or amidine carbon atoms): A 171.8, 169.9, 156.8

Eksempel 8 Example 8

Parallel Syntese av Alkoksvamidiner Parallel Synthesis of Alkoxavamidines

Denne syntesen ble utført i en 96-brønn Robbins blokkerer. Til brønner inneholdende en passende mengde av O-substituert hydroksylamin (spesifisert nedenfor; idet alle er kommersielt tilgjengelig eller ble fremstilt ved anvendelse av velkjente litteratur prosedyrer) ble tilsatt en løsning av Ph(3-Cl)(5-OCF3)-(Æ)CH(OH)C(0)-Aze-Pab(Teoc) This synthesis was performed in a 96-well Robbins blocker. To wells containing an appropriate amount of O-substituted hydroxylamine (specified below; all being commercially available or prepared using well-known literature procedures) was added a solution of Ph(3-Cl)(5-OCF3)-(Æ) CH(OH)C(0)-Aze-Pab(Teoc)

(10 mg; 17 umol; se Eksempel 6(x) ovenfor) i acetonitril (1,0 ml). Blokkeringsmidlet ble forseglet og reaksjonsblandingen ble rotert natten over i en ovn ved 60°C. Etter avkjøling og filtrering, ble de faste stoffene vasket med acetonitril (3 x 0,3 ml). De samlede (10 mg; 17 µmol; see Example 6(x) above) in acetonitrile (1.0 mL). The blocking agent was sealed and the reaction mixture was rotated overnight in an oven at 60°C. After cooling and filtration, the solids were washed with acetonitrile (3 x 0.3 mL). They collected

væskefraksjoner ble konsentrert i en vakuumsentrifuge. Residuet ble fordelt mellom vann (0,4 ml) og etylacetat (0,4 ml). Etter at væske-væske ekstraksjonen var avsluttet, ble alt filtrert gjennom en kolonne av Hydromatriks™. Etter vasking tre ganger med etylacetat, ble de samlede filtrater konsentrert i en vakuumsentrifuge. Avbeskyttelse ble utført ved tilsetning av metylenklorid (0,1 ml) og trifluroeddiksyre (0,3 ml). Etter omrøring ved romtemperatur i 3 timer, ble løsningsmidlene fjernet i vakuum. Residuet ble fordelt mellom vandig mettet natriumhydrogenkarbonat (0,5 ml) og etylacetat (0,5 ml). Etter ekstraksjon, filtrering gjennom Hydromatriks og konsentrasjon ( vid infra) ble residuet oppløst i isopropanol/vann (7/3) (1 ml). Omtrent 2% av denne løsningen ble fjernet og fortynnet med isopropanol/vann (7/3) (1 ml) for LC-MS analyse. Etter fjerning av løsningsmidlene i vakuum ble det faste residuet overført til en 96-brønn plate ved anvendelse av acetonitril og etylacetat for å oppløse forbindelsen. Løsningsmidlene ble avdampet i en vakuum sentrifuge, hvilket gir de følgende tittelforbindelser: liquid fractions were concentrated in a vacuum centrifuge. The residue was partitioned between water (0.4 ml) and ethyl acetate (0.4 ml). After the liquid-liquid extraction was finished, everything was filtered through a column of Hydromatriks™. After washing three times with ethyl acetate, the combined filtrates were concentrated in a vacuum centrifuge. Deprotection was carried out by adding methylene chloride (0.1 ml) and trifluoroacetic acid (0.3 ml). After stirring at room temperature for 3 hours, the solvents were removed in vacuo. The residue was partitioned between aqueous saturated sodium bicarbonate (0.5 mL) and ethyl acetate (0.5 mL). After extraction, filtration through Hydromatrix and concentration (vid infra), the residue was dissolved in isopropanol/water (7/3) (1 ml). About 2% of this solution was removed and diluted with isopropanol/water (7/3) (1 mL) for LC-MS analysis. After removal of the solvents in vacuo, the solid residue was transferred to a 96-well plate using acetonitrile and ethyl acetate to dissolve the compound. The solvents were evaporated in a vacuum centrifuge, giving the following title compounds:

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OCH2-3-(5-Me-isoksazol)) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OCH2-3-(5-Me-isoxazole))

(fra 3-[(aminooksy)metyl]-5-metylisoksazol x HC1 (18 mg; 0,11 mmol)). Utbytte: 3,64 mg (35%) (MS (m/z) 596 (M + 1)<+>); (from 3-[(aminooxy)methyl]-5-methylisoxazole x HCl (18 mg; 0.11 mmol)). Yield: 3.64 mg (35%) (MS (m/z) 596 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OCH2-3-pyridin) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OCH2-3-pyridine)

(fra 3-[(aminooksy)metyl]pyridin x 2 HC1 (19 mg; 96 umol). Utbytte: 5,14 mg (50%) (MS (m/z) 592 (M + 1)<+>); (from 3-[(aminooxy)methyl]pyridine x 2 HCl (19 mg; 96 µmol). Yield: 5.14 mg (50%) (MS (m/z) 592 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(0/Bu) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(0/Bu)

(fra Ø-isobutyl hydroksylamin x HC1 (17 mg; 140 umol). Utbytte: 4,4 mg (45%). MS (m/z) 557 (M + 1)<+>); (from Ø-isobutyl hydroxylamine x HCl (17 mg; 140 umol). Yield: 4.4 mg (45%). MS (m/z) 557 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OEt) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OEt)

(fra O-etyl hydroksylamin x HC1 (14 mg; 140 umol). Utbytte: 4,04 mg (42%). MS (m/z) 529 (M+l)<+>); (from O-ethyl hydroxylamine x HCl (14 mg; 140 µmol). Yield: 4.04 mg (42%. MS (m/z) 529 (M+1)<+>);

Ph(3-Cl)(5-OCF3)-(Æ)CH(OH)C(0)-Aze-Pab(OBn) Ph(3-Cl)(5-OCF3)-(Æ)CH(OH)C(0)-Aze-Pab(OBn)

(fra Ø-benzylhydroksylamin x HC1 (17 mg; 110 umol). Utbytte: 3,22 mg (29%). MS (m/z) 591 (M + 1)<+>); (from β-benzylhydroxylamine x HCl (17 mg; 110 umol). Yield: 3.22 mg (29%). MS (m/z) 591 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OcHeksyl) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OcHexyl)

(fra Ø-cykloheksyl hydroksylamin x HC1 (15 mg; 99 umol). Utbytte: 2,9 mg (26%). MS (m/z) 583 (M + 1)<+>); (from Ø-cyclohexyl hydroxylamine x HCl (15 mg; 99 umol). Yield: 2.9 mg (26%). MS (m/z) 583 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(Æ)CH(OH)C(0)-Aze-Pab(OcBu) Ph(3-Cl)(5-OCF3)-(Æ)CH(OH)C(0)-Aze-Pab(OcBu)

(fra Ø-cyklobutyl hydroksylamin x HC1 (17 mg; 140 umol). Utbytte: 3,3 mg (30%). MS (m/z) 555 (M + 1)<+>); (from Ø-cyclobutyl hydroxylamine x HCl (17 mg; 140 umol). Yield: 3.3 mg (30%). MS (m/z) 555 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OCH2CH20Ph(3-CF3)) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OCH2CH20Ph(3-CF3))

(fra 0-[2-[3-(trifluormetyl)fenoksy]etyl]hydroksylamin x HC1 (24 mg; 93 umol). Utbytte: 6,52 mg (46%). MS (m/z) 689 (M+l)<+>); (from 0-[2-[3-(trifluoromethyl)phenoxy]ethyl]hydroxylamine x HCl (24 mg; 93 µmol). Yield: 6.52 mg (46%). MS (m/z) 689 (M+l )<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OBn(4-Cl)) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OBn(4-Cl))

(fra 0-(4-klorbenzyl)hydroksylamin x HC1 (16 mg; 82 umol). Utbytte: 3,47 mg (29%). MS (m/z) 625 (M + 1)<+>); (from 0-(4-chlorobenzyl)hydroxylamine x HCl (16 mg; 82 µmol). Yield: 3.47 mg (29%). MS (m/z) 625 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(/2)CH(OH)C(0)-Aze-Pab(OBn(3-MeO)) Ph(3-Cl)(5-OCF3)-(/2)CH(OH)C(0)-Aze-Pab(OBn(3-MeO))

(fra 0-(3-metoksybenzyl)hydroksylamin x HC1 (18 mg; 94 umol). Utbytte: 4,33 mg (36%). MS (m/z) 621 (M+l)<+>); (from 0-(3-methoxybenzyl)hydroxylamine x HCl (18 mg; 94 µmol). Yield: 4.33 mg (36%). MS (m/z) 621 (M+1)<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OBn(2-Br)) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OBn(2-Br))

(fra 0-(2-brombenzyl)hydroksylamin x HC1 (23 mg; 96 umol). Utbytte: 3,87 mg (30%). MS (m/z) 671 (M + 1)<+>); (from 0-(2-bromobenzyl)hydroxylamine x HCl (23 mg; 96 µmol). Yield: 3.87 mg (30%). MS (m/z) 671 (M + 1)<+>);

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OBn(4-Me)) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(OBn(4-Me))

(fra 0-(4-metylbenzyl)hydroksylamin x HC1 (14 mg; 81umol). Utbytte: 2,91 mg (25%). (from 0-(4-methylbenzyl)hydroxylamine x HCl (14 mg; 81 µmol). Yield: 2.91 mg (25%).

MS (m/z) 605 (M + 1)<+>); og MS (m/z) 605 (M + 1)<+>); and

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(0-4-heptyl) Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab(0-4-heptyl)

(fra 0-(4-heptyl)hydxoksylamin x HC1 (15 mg; 89 umol). Utbytte: 17 mg (100%). MS (m/z) 599 (M + 1)<+>). (from 0-(4-heptyl)hydoxylamine x HC1 (15 mg; 89 µmol). Yield: 17 mg (100%). MS (m/z) 599 (M + 1)<+>).

Eksempel 9 Example 9

Ph( 3- Cn( 5- OCHF2)-(^ CH( CH7QH) C( 0)- Aze- Pab x HOAc Ph( 3- Cn( 5- OCHF2)-(^ CH( CH7QH) C( 0)- Aze- Pab x HOAc

(i) 3- klor- 5- metoksvbenzosvre (i) 3-chloro-5-methoxybenzoic acid

Magnesiumspon (Fluka purum for Grignard reaksjoner) ble forbehandlet på følgende måte: sponet ble plassert i en glass sinteret trakt og 0,1 M av saltsyre ble hellet i dem. Sponene ble omrørt med en glass stav i noen få sekunder og deretter ble syren vasket bort med 3 porsjoner vann. Til slutt, ble sponet vasket med 2 porsjoner av aceton og satt på flasker. Tetrahydrofuran (100 ml, 99,95%) ble tørket ved tilsetning av RedAl (1 g, 70% vekt. i toluen). Forbehandlet magnesiumspon (5 g, 200 mmol) ble plassert i en rundbunnet kolbe og ble spylt med nitrogen 3 ganger. Dikloranisol (26 g, 146 mmol) ble oppløst i THF (100 ml, RedAl-tørket) og dibrometan (1,8 g, Magnesium shavings (Fluka purum for Grignard reactions) were pretreated as follows: the shavings were placed in a glass sintered funnel and 0.1 M of hydrochloric acid was poured into them. The chips were stirred with a glass rod for a few seconds and then the acid was washed away with 3 portions of water. Finally, the chips were washed with 2 portions of acetone and put into bottles. Tetrahydrofuran (100 mL, 99.95%) was dried by addition of RedAl (1 g, 70% wt. in toluene). Pretreated magnesium shavings (5 g, 200 mmol) were placed in a round bottom flask and flushed with nitrogen 3 times. Dichloroanisole (26 g, 146 mmol) was dissolved in THF (100 mL, RedAl-dried) and dibromoethane (1.8 g,

10 mmol) ble tilsatt. Reaksjonsblandingen ble spylt med nitrogen og deretter tilbakeløpskokt i 2 timer. Oppvarmning ble avbrudt og tørris (10 g) ble tilsatt porsjons vis over 2 minutter. Når all tørris var oppløst, ble reaksjonsblandingen hellet i is inneholdende saltsyre 10 mmol) was added. The reaction mixture was flushed with nitrogen and then refluxed for 2 hours. Heating was interrupted and dry ice (10 g) was added in portions over 2 minutes. When all the dry ice had dissolved, the reaction mixture was poured into ice containing hydrochloric acid

(400 ml, 2 M). Ekstra aktiv opparbeiding (eter, 300 ml) ga 11,2 g, (400ml, 2M). Extra active work-up (ether, 300 ml) gave 11.2 g,

60,2 mmol (utbytte: 41%) av sub-tittelforbindelsen. 60.2 mmol (yield: 41%) of the sub-title compound.

■H-NMR (500 MHz; aceton-d6): A 7,57 (m, 1H), 7,49 (m, 1H), 7,23 (m, 1H), 3,91 (s, 3H) ■H-NMR (500 MHz; acetone-d6): A 7.57 (m, 1H), 7.49 (m, 1H), 7.23 (m, 1H), 3.91 (s, 3H)

(ii) 3- klor- 5- hvdroksvbenzosvre (ii) 3-chloro-5-hydroxybenzoic acid

Alumina (1,65 g, 60 mmol) og jod (21 g, 82 mmol) ble tilbakeløpskokt i toluen (200 ml) i 2 timer. Deretter, ble 3-klor-5-metoksybenzosyre Alumina (1.65 g, 60 mmol) and iodine (21 g, 82 mmol) were refluxed in toluene (200 mL) for 2 h. Then, was 3-chloro-5-methoxybenzoic acid

(11,2 g, 60,2 mmol; se trinn (i) ovenfor) oppløst i toluen (50 ml) tilsatt, sammen med tetrabutylammoniumjodid (1,5 g, 4 mmol) og blandingen ble tilbakeløpskokt i ytterligere 2 timer. Etter avkjøling til omgivelsestemperatur, ga ekstra aktiv opparbeiding 8,7 g, 50 mmol (utbytte: 83%) av sub-tittelforbindelse. (11.2 g, 60.2 mmol; see step (i) above) dissolved in toluene (50 mL) was added, along with tetrabutylammonium iodide (1.5 g, 4 mmol) and the mixture was refluxed for a further 2 h. After cooling to ambient temperature, additional active workup afforded 8.7 g, 50 mmol (yield: 83%) of sub-title compound.

'H-NMR (300 MHz; aceton-de): A 9,27 (s, 1H), 7,48 (m, 1H), 7,44 (m, 1H), 7,11 (m, 1H) 1H-NMR (300 MHz; acetone-de): Δ 9.27 (s, 1H), 7.48 (m, 1H), 7.44 (m, 1H), 7.11 (m, 1H)

(iii) 3- klor- 5- difluormetoksvbenzosvre (iii) 3-chloro-5-difluoromethoxybenzoic acid

3-klor-5-hydroksybenzosyre (6,4 g, 37,2 mmol; se trinn (ii) ovenfor) oppløst i kloroform (200 ml) ble overført til en 500 ml trehalset rundbunnet kolbe utstyrt med en tørris kondenser og et gass innførselsrør. Natriumhydroksyd (100 ml, 5 M) ble tilsatt med kraftig omrøring. Klordifluormetan (Freon 22; 25 g, 290 mmol) ble tilsatt porsjonsvis gjennom gass innførselsrøret ved omgivelsestemperatur. Etter 2 timer, var reaksjonen fullstendig. Ekstra aktiv opparbeiding ga 6,2 g, 28 mmol (utbytte: 75%) av sub-tittelforbindelse. 3-Chloro-5-hydroxybenzoic acid (6.4 g, 37.2 mmol; see step (ii) above) dissolved in chloroform (200 mL) was transferred to a 500 mL three-necked round-bottom flask equipped with a dry ice condenser and a gas inlet tube . Sodium hydroxide (100 mL, 5 M) was added with vigorous stirring. Chlorodifluoromethane (Freon 22; 25 g, 290 mmol) was added portionwise through the gas inlet tube at ambient temperature. After 2 hours, the reaction was complete. Additional active workup afforded 6.2 g, 28 mmol (yield: 75%) of sub-title compound.

'H-NMR (500 MHz; aceton-de): A 7,87 (m, 1H), 7,74 (m, 1H), 7,54 (m, 1H), 7,19 (t, 1H, Jh-f73Hz) 1H-NMR (500 MHz; acetone-de): Δ 7.87 (m, 1H), 7.74 (m, 1H), 7.54 (m, 1H), 7.19 (t, 1H, Jh -f73Hz)

(iv) 3- klor- 5- difluormetoksv- N- metoksv- N- metvlbenzamid 3-klor-5-difluormetoksybenzosyre (1,8 g, 8 mmol; se trinn (iii) ovenfor) og oksalylklorid (1,5 g,ll,8 mmol) ble oppløst i metylenklorid (50 ml). DMF (2 dråper) ble tilsatt og reaksjonsblandingen ble omrørt ved omgivelsestemperatur i 30 minutter. Deretter, ble A^,0-dimetylhydroksylamin (1 g, 10,2 mmol) og trietylamin (3 g, 30 mmol) tilsatt og etter ytterligere 10 minutter omrøring ved omgivelsestemperatur, ble reaksjonsblandingen konsentrert ved redusert trykk. Residuet ble tatt opp i eter (100 ml) og vann (50 ml). Etter separering, ble den organiske fasen vasket med saltvann, tørket over natriumsulfat, filtrert og konsentrert. Dette residuet ble kromatografert på silika (heksan/etylacetat 2:1) som ga 2 g, 7,5 mmol (93%) av sub-tittelforbindelse. (iv) 3-chloro-5-difluoromethoxy-N-methoxy-N-methylbenzamide 3-chloro-5-difluoromethoxybenzoic acid (1.8 g, 8 mmol; see step (iii) above) and oxalyl chloride (1.5 g, II .8 mmol) was dissolved in methylene chloride (50 ml). DMF (2 drops) was added and the reaction mixture was stirred at ambient temperature for 30 minutes. Then, N , O -dimethylhydroxylamine (1 g, 10.2 mmol) and triethylamine (3 g, 30 mmol) were added and after a further 10 minutes of stirring at ambient temperature, the reaction mixture was concentrated under reduced pressure. The residue was taken up in ether (100 ml) and water (50 ml). After separation, the organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated. This residue was chromatographed on silica (hexane/ethyl acetate 2:1) to give 2 g, 7.5 mmol (93%) of sub-title compound.

'H-NMR (400 MHz; CDC13): A 7,54 (m, 1H), 7,37 (m, 1H), 7,27 (m, 1H), 6,53 (t, 1H, JH-f 73 Hz) 1H-NMR (400 MHz; CDCl 3 ): Δ 7.54 (m, 1H), 7.37 (m, 1H), 7.27 (m, 1H), 6.53 (t, 1H, JH-f 73Hz)

(v) 3- klor- 5- difluormetoksvacetofenon (v) 3-chloro-5-difluoromethoxyacetophenone

3-klor-5-difluormetoksy-N-metoksy-N-metylbenzamid (2 g, 3-chloro-5-difluoromethoxy-N-methoxy-N-methylbenzamide (2 g,

7,5 mmol; se trinn (iv) ovenfor) ble oppløst i eter (100 ml) og avkjølt under nitrogen til - 70°C. Metyllitium (7 ml, 11 mmol, 1,6 M i eter) ble tilsatt dråpevis med en sprøyte til den omrørte reaksjonsblanding over 1 minutt. Tørris-badet ble fjernet og blandingen fikk nå omgivelsestemperatur før reaksjonen ble stanset med ammoniumklorid-løsning (50 ml, 5% NH4CI i vann). Den organiske fasen ble vasket med saltvann, tørket over natriumsulfat, filtrert og konsentrert ved redusert trykk. Residuet ble kromatografert på silika (heksametylacetat 2:1) som ga 1,5 g, 6,8 mmol (utbytte: 90%) av sub-tittelforbindelse. 7.5 mmol; see step (iv) above) was dissolved in ether (100 mL) and cooled under nitrogen to -70°C. Methyllithium (7 mL, 11 mmol, 1.6 M in ether) was added dropwise via syringe to the stirred reaction mixture over 1 minute. The dry ice bath was removed and the mixture allowed to reach ambient temperature before the reaction was quenched with ammonium chloride solution (50 mL, 5% NH 4 Cl in water). The organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica (hexamethyl acetate 2:1) to give 1.5 g, 6.8 mmol (yield: 90%) of sub-title compound.

•H-NMR (600 MHz; CDC13): A 7,77 (m, 1H), 7,59 (m, 1H), 7,35 (m, 1H), 6,56 (t, 1H, JH-f 73 Hz), 2,60 (s, 3H) H-NMR (600 MHz; CDCl 3 ): A 7.77 (m, 1H), 7.59 (m, 1H), 7.35 (m, 1H), 6.56 (t, 1H, JH-f 73 Hz), 2.60 (s, 3H)

(vi) 3- klor- 5- difluormetoksvfenvleddiksyre- metvlester (vi) 3-Chloro-5-difluoromethoxysulfenvacetic acid methyl ester

3-klor-5-difluormetoksyacetofenon (1,5 g, 6,8 mmol; se trinn (v) ovenfor) ble oppløst i metylenklorid (200 ml). Thallium(III) nitrat x 3MeOH på K-10 montmorillonite (6 g, 10 mmol (ca 0,6 mmol/g); se J. Am. Chem. Soc, 98,6750 (1976)) ble tilsatt og blandingen ble omrørt ved omgivelsestemperatur i 20 timer. Blandingen ble filtrert og filtratet ble vasket med natriumbikarbonat (100 ml, 0,5 M), tørket over natriumsulfat, filtrert og konsentrert ved redusert trykk. Residuet ble kromatografert på silika (heksan/etylacetat 2:1) som ga 1 3-Chloro-5-difluoromethoxyacetophenone (1.5 g, 6.8 mmol; see step (v) above) was dissolved in methylene chloride (200 mL). Thallium(III) nitrate x 3MeOH on K-10 montmorillonite (6 g, 10 mmol (ca. 0.6 mmol/g); see J. Am. Chem. Soc, 98, 6750 (1976)) was added and the mixture was stirred at ambient temperature for 20 hours. The mixture was filtered and the filtrate was washed with sodium bicarbonate (100 mL, 0.5 M), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica (hexane/ethyl acetate 2:1) which gave 1

g. g.

4 mmol (utbytte: 56%) av sub-tittelforbindelse. 4 mmol (yield: 56%) of sub-title compound.

^-NMR (500 MHz; CDC13): A 7,14 (m, 1H), 7,06 (m, 1H), 6,96 (m, 1H), 6,50 (t, 1H, JH-f 3-NMR (500 MHz; CDCl 3 ): A 7.14 (m, 1H), 7.06 (m, 1H), 6.96 (m, 1H), 6.50 (t, 1H, JH-f

73 Hz), 3,72 (s, 3H), 3,60 (s,lH) (vii) a- Formyl( 3- klor- 5- difluormetoksyfenvl) eddiksvre- metylester 3-klor-5-difluormetoksyfenyleddiksyre-metylester (1 g, 4 mmol; se trinn (vi) ovenfor) og metylformiat (1 g, 16 mmol) ble oppløst i eter (100 ml) og avkjølt i et isbad (ca. 2°C). Deretter, ble finfordelt kuttet natrium (180 mg, 7,8 mmol) og metanol (1 ml) tilsatt og blandingen ble latt stå i is-bad med omrøring natten over. Vann (100 ml) ble forsiktig tilsatt og fasene ble separert. Den vannet inneholdende fasen ble surgjort med saltsyre (2 M) til pH 1 og ekstrahert med eter (2 x 100 ml). Ekstrakten ble tørket over natriumsulfat, filtrert og konsentrert ved redusert trykk. Residuet ble kromatografert på silika (heksametylacetat (1:1)) som ga 400 mg, 1,4 mmol (utbytte: 36%) av sub-tittelforbindelse. 73 Hz), 3.72 (s, 3H), 3.60 (s,lH) (vii) a- Formyl (3-chloro-5-difluoromethoxyphenyl) acetic acid methyl ester 3-chloro-5-difluoromethoxyphenylacetic acid methyl ester (1 g, 4 mmol; see step (vi) above) and methyl formate (1 g, 16 mmol) were dissolved in ether (100 mL) and cooled in an ice bath (ca. 2°C). Then, finely divided cut sodium (180 mg, 7.8 mmol) and methanol (1 mL) were added and the mixture was left in an ice bath with stirring overnight. Water (100 mL) was carefully added and the phases were separated. The aqueous phase was acidified with hydrochloric acid (2 M) to pH 1 and extracted with ether (2 x 100 mL). The extract was dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed on silica (hexamethyl acetate (1:1)) to give 400 mg, 1.4 mmol (yield: 36%) of sub-title compound.

'H-NMR (400 MHz): A 12,10 (d, 1H), 7,32 (d, 1H), 7,11 (m, 1H), 7,07 (m, 1H), 6,94 (m, 1H), 6,51 (t, 1H, JF.H 73), 3,83 (s, 3H) 1H NMR (400 MHz): Δ 12.10 (d, 1H), 7.32 (d, 1H), 7.11 (m, 1H), 7.07 (m, 1H), 6.94 ( m, 1H), 6.51 (t, 1H, JF.H 73), 3.83 (s, 3H)

(viii) 3- klor- 5- difluormetoksvtropic syre (viii) 3-chloro-5-difluoromethoxystropic acid

a-Formyl(3-klor-5-difluormetoksyfenyl)eddiksyre-metylester α-Formyl (3-chloro-5-difluoromethoxyphenyl)acetic acid methyl ester

(400 mg, 1,4 mmol; se trinn (vii) ovenfor) ble oppløst i THF:metanol (50 ml, 9:1). Natrium-borhydrid ble tilsatt og blandingen ble omrørt ved omgivelsestemperatur i 30 minutter. Vann ble tilsatt og blandingen ble konsentrert for å produsere en vandig suspensjon, som ble tatt opp i etylacetat og vann. Fasene ble separert og den organiske fasen ble vasket med natriumklorid (15% i vann), tørket over natriumsulfat, filtrert og konsentrert ved redusert trykk. Residuet ble oppløst i metanol (30 ml) og hydrolysen med natriumhydroksyd (400 mg, 1.4 mmol; see step (vii) above) was dissolved in THF:methanol (50 mL, 9:1). Sodium borohydride was added and the mixture was stirred at ambient temperature for 30 minutes. Water was added and the mixture was concentrated to produce an aqueous suspension, which was taken up in ethyl acetate and water. The phases were separated and the organic phase was washed with sodium chloride (15% in water), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was dissolved in methanol (30 ml) and hydrolysed with sodium hydroxide

(1 ml, 10 M) ved omgivelsestemperatur i 10 minutter. Ekstraktiv opparbeiding ga 180 mg, 0,68 mmol (utbytte: 48%) av sub-tittelforbindelse. (1 ml, 10 M) at ambient temperature for 10 minutes. Extractive work-up gave 180 mg, 0.68 mmol (yield: 48%) of sub-title compound.

'H-NMR (500 MHz; CDC13): A 7,18 (m, 1H), 7,10 (m, 1H), 7,00 (m. 1H), 6,50 (t, 1H, JF.H 73), 4,11 (m, 1H), 3,90 (m, 1H), 3,84 (m, 1H) 1H NMR (500 MHz; CDCl 3 ): Δ 7.18 (m, 1H), 7.10 (m, 1H), 7.00 (m. 1H), 6.50 (t, 1H, JF.H 73), 4.11 (m, 1H), 3.90 (m, 1H), 3.84 (m, 1H)

(ix) Ph( 3- Cl)( 5- OCHFz)-( 5) CH( CH20H) C( 0)- Azé- Pab x HOAc (ix) Ph( 3- Cl)( 5- OCHFz)-( 5) CH( CH2OH) C( 0)- Azé- Pab x HOAc

3-klor-5-difluormetoksytropinsyre (180 mg, 0,7 mmol; se trinn (viii) ovenfor), H-Aze-Pab(Teoc) x HC1 (450 mg, 1 mmol) og PyBOP (530 mg, 1 mmol) ble oppløst i DMF (10 ml), hvoretter DIPEA (550 mg, 3,9 mmol) ble tilsatt. Blandingen ble omrørt ved omgivelsestemperatur i 1 time før den ble fortynnet med saltvann (20 ml, 15% NaCl) og ekstrahert med etylacetat (40 ml). Ekstrakten ble tørket over natriumsulfat, filtrert og inndampet til tørrhet. Residuet ble oppløst i metylenklorid (5 ml) og trifluroeddiksyre (5 ml) ble tilsatt. Etter 1 time ved omgivelsestemperatur, ble blandingen av diastereomerene inndampet til tørrhet og residuet ble kromatografert på en revers fase kolonne (acetonitril:vann (30:70), buffer: ammoniumacetat 0,1 M). Frysetørking ga 36 mg, 0,067 mmol (utbytte: 10,4%) av tittelforbindelsen. 3-Chloro-5-difluoromethoxytropic acid (180 mg, 0.7 mmol; see step (viii) above), H-Aze-Pab(Teoc) x HC1 (450 mg, 1 mmol) and PyBOP (530 mg, 1 mmol) was dissolved in DMF (10 mL), after which DIPEA (550 mg, 3.9 mmol) was added. The mixture was stirred at ambient temperature for 1 hour before being diluted with brine (20 mL, 15% NaCl) and extracted with ethyl acetate (40 mL). The extract was dried over sodium sulfate, filtered and evaporated to dryness. The residue was dissolved in methylene chloride (5 ml) and trifluoroacetic acid (5 ml) was added. After 1 hour at ambient temperature, the mixture of diastereomers was evaporated to dryness and the residue was chromatographed on a reverse phase column (acetonitrile:water (30:70), buffer: ammonium acetate 0.1 M). Freeze drying gave 36 mg, 0.067 mmol (yield: 10.4%) of the title compound.

MS (ES) 481(M + 1)<+>MS (ES) 481(M + 1)<+>

•H-NMR (400 MHz; CDCI3): A 7,77 (d, 2H), 7,57 (d, 2H), 7,30 (m, 1H), 7,13 (m, 2H), 6,87 (t, 1H, JF.H 73 Hz), 4,76 (m, 1H), 4,55 (s, 2H), 4,37 (m, 1H), 4,03 (m, 2H), 3,82 (m, 1H), 3,72 (m, 1H), 2,53 (m, 1H), 2,28 (m, 1H), 1,92 (s, 1,5H) H-NMR (400 MHz; CDCl 3 ): A 7.77 (d, 2H), 7.57 (d, 2H), 7.30 (m, 1H), 7.13 (m, 2H), 6, 87 (t, 1H, JF.H 73 Hz), 4.76 (m, 1H), 4.55 (s, 2H), 4.37 (m, 1H), 4.03 (m, 2H), 3 .82 (m, 1H), 3.72 (m, 1H), 2.53 (m, 1H), 2.28 (m, 1H), 1.92 (s, 1.5H)

<l3>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) A 172,3,171,9,167,2 <l3>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms) A 172,3,171,9,167,2

Eksempel 10 Example 10

Ph( 3- Cl)( 5- OCFO-( S) CH( CH7OH) C( Q)- Aze- Pab x TFA Ph( 3- Cl)( 5- OCFO-( S) CH( CH7OH) C( Q)- Aze- Pab x TFA

(i) 3- klor- 5- trifluormetoksvbenzvlmesylat (i) 3-Chloro-5-trifluoromethoxybenzoyl mesylate

Til en løsning av 3-klor-5-trifluormetoksybenzylalkohol (6,1 g, To a solution of 3-chloro-5-trifluoromethoxybenzyl alcohol (6.1 g,

26,9 mmol; se Eksempel 6(v) ovenfor) i CH2CI2 (250 ml) ved 0°C under en nitrogen- 26.9 mmol; see Example 6(v) above) in CH2Cl2 (250 mL) at 0°C under a nitrogen

atmosfære ble tilsatt DIPEA (4,2 g, 32,3 mmol) og metansulfonylklorid (3,4 g, 29,6 atmosphere was added DIPEA (4.2 g, 32.3 mmol) and methanesulfonyl chloride (3.4 g, 29.6

mmol). Løsningen ble omrørt ved 0°C i 1,5 timer og behandlet med H2O. De organiske faser ble separert og deretter vasket med H2O (lx), IN HC1 (lx), H2O (lx) og vandig NaHC03 (lx) og deretter tørket (Na2S04), filtrert og konsentrert, hvilket gir sub-tittelforbindelsen (8,2 g, 99%) som en olje. mmol). The solution was stirred at 0°C for 1.5 hours and treated with H 2 O. The organic phases were separated and then washed with H 2 O (lx), IN HCl (lx), H 2 O (lx) and aqueous NaHCO 3 (lx) and then dried (Na 2 SO 4 ), filtered and concentrated to give the sub-title compound (8.2 g, 99%) as an oil.

<*>H NMR (300 MHz, CDC13): A 7,37 (s, 1H), 7,28 (s, 1H), 7,18 (s, 1H) 5,23 (s, 2H), 3,07 <*>H NMR (300 MHz, CDCl 3 ): Δ 7.37 (s, 1H), 7.28 (s, 1H), 7.18 (s, 1H) 5.23 (s, 2H), 3, 07

(s, 3H) (p, 3H)

(ii) 3- klor- 5- trifluormetoksvbenzylcvanid (ii) 3-Chloro-5-trifluoromethoxybenzyl cyanide

Til en løsning av 3-klor-5-trifluormetoksybenzylmesylat (8,2 g, To a solution of 3-chloro-5-trifluoromethoxybenzyl mesylate (8.2 g,

26,8 mmol; se trinn (i) ovenfor) i DMSO (50 ml) ble tilsatt natriumcyanid (2,6 g, 53,6 mmol). Den resulterende heterogene løsning ble oppvarmet til 50°C og ultralydbehandlet i 1 time. Reaksjonsblandingen ble avkjølt og fordelt mellom Et20 og H20. De organiske faser ble vasket med H20 (2x) og saltvann (2x). Den samlede vandige faser ble ekstrahert med Et20 (lx). De samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert under en lav varme og partielt vakuum, hvilket gir sub-tittelforbindelsen (6,3 g, 100%) som en rødaktig flyktig olje som ble anvendt i neste trinn uten ytterligere rensning. 26.8 mmol; see step (i) above) in DMSO (50 mL) was added sodium cyanide (2.6 g, 53.6 mmol). The resulting heterogeneous solution was heated to 50°C and sonicated for 1 hour. The reaction mixture was cooled and partitioned between Et 2 O and H 2 O. The organic phases were washed with H 2 O (2x) and brine (2x). The combined aqueous phases were extracted with Et 2 O (1x). The combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated under low heat and partial vacuum to give the sub-title compound (6.3 g, 100%) as a reddish volatile oil which was used in the next step without further purification.

<*>H NMR (300 MHz, CDCI3) A 7,32 (s, 1H), 7,24 (s, 1H), 7,12 (s, 1H), 3,78 (s, 2H) <*>H NMR (300 MHz, CDCl 3 ) Δ 7.32 (s, 1H), 7.24 (s, 1H), 7.12 (s, 1H), 3.78 (s, 2H)

(iii) 3- klor- 5- trifluormetoksvfenvleddiksvre (iii) 3-chloro-5-trifluoromethoxysulfenylacetic acid

Til en løsning av 3-klor-5-trifluormetoksybenzylcyanid (6,3 g, To a solution of 3-chloro-5-trifluoromethoxybenzyl cyanide (6.3 g,

26,7 mmol; se trinn (ii) ovenfor) i 2-propanol (100 ml) ble det tilsatt vann (200 ml) og kaliumhydroksyd (7,5 g, 133,5 mmol). Løsningen ble tilbakeløpskokt i 18 timer, avkjølt til romtemperatur og 2-propanol ble fjernet i vakuum. Den vandige fasen ble vasket med CH2C12 (2x) og vaskevæskene kastet. Den basiske vandige fasen ble surgjort med 2N HC1 og ekstrahert med CH2C12 (3x). CH2C12 ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelsen (5,2 g, 76%) som en olje som ble anvendt i neste trinn uten ytterligere rensning. 26.7 mmol; see step (ii) above) in 2-propanol (100 ml) was added water (200 ml) and potassium hydroxide (7.5 g, 133.5 mmol). The solution was refluxed for 18 hours, cooled to room temperature and 2-propanol was removed in vacuo. The aqueous phase was washed with CH 2 Cl 2 (2x) and the washings discarded. The basic aqueous phase was acidified with 2N HCl and extracted with CH 2 Cl 2 (3x). CH 2 Cl 2 extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (5.2 g, 76%) as an oil which was used in the next step without further purification.

'H NMR (300 MHz, CDCI3): A 7,25 (s, 1H), 7,19 (s, 1H), 7,08 (s, 1H), 3,68 (s, 2H) 1 H NMR (300 MHz, CDCl 3 ): Δ 7.25 (s, 1H), 7.19 (s, 1H), 7.08 (s, 1H), 3.68 (s, 2H)

(iv) Etyl 3- klor- 5- trifluormetoksvfenvlacetat (iv) Ethyl 3-chloro-5-trifluoromethoxysulfonyl acetate

Til en løsning av 3-klor-5-trifluormetoksyfenyleddiksyre (5,2 g, To a solution of 3-chloro-5-trifluoromethoxyphenylacetic acid (5.2 g,

20,4 mmol; se trinn (iii) ovenfor) i EtOH (600 ml) ble det tilsatt svovelsyre (mange dråper). Løsningen ble tilbakeløpskokt i 18 timer, avkjølt til romtemperatur, nøytralisert med fast stoff NaHC03 og EtOH fjernet i vakuum. Residuet ble fortynnet med EtOAc deretter vasket med H20 (lx), vandig NaHC03 (lx) og saltvann (lx). De organiske faser 20.4 mmol; see step (iii) above) in EtOH (600 mL) was added sulfuric acid (many drops). The solution was refluxed for 18 hours, cooled to room temperature, neutralized with solid NaHCO 3 and EtOH removed in vacuo. The residue was diluted with EtOAc then washed with H 2 O (lx), aqueous NaHCO 3 (lx) and brine (lx). The organic phases

ble tørket (Na2SC«4), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelsen (5,5 g, 96%) som en olje som ble anvendt i neste trinn uten ytterligere rensning. was dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give the sub-title compound (5.5 g, 96%) as an oil which was used in the next step without further purification.

'H NMR (300 MHz, CDC13) A 7,24 (s, 1H), 7,16 (s, 1H), 7,07 (s, 1H), 4,13-4,22 (q, 7 = 8 Hz, 2H), 3,63 (s, 2H), 1,24-1,32 (t, 7 = 8 Hz, 3H) 1 H NMR (300 MHz, CDCl 3 ) Δ 7.24 (s, 1H), 7.16 (s, 1H), 7.07 (s, 1H), 4.13-4.22 (q, 7 = 8 Hz, 2H), 3.63 (s, 2H), 1.24-1.32 (t, 7 = 8 Hz, 3H)

(v) Ph( 3- Cl)( 5- OCFi)-(/ ?. S) CH( CHO) C( Q) OEt (v) Ph( 3- Cl)( 5- OCFi)-(/ ?. S) CH( CHO) C( Q) OEt

Til en løsning av etyl 3-klor-5-trifluormetoksyfenylacetat (4,5 g, To a solution of ethyl 3-chloro-5-trifluoromethoxyphenyl acetate (4.5 g,

15,9 mmol; se trinn (iv) ovenfor) i vannfri THF (400 ml) under en nitrogen-atmosfære ved mindre enn 0°C (is-MeOH bad) ble det tilsatt natriumetoksyd (4,5 g, 63,6 mmol). Den kalde løsningen ble omrørt i 40 minutter og etylformiat (8,1 g, 111,3 mmol) ble tilsatt. Løsningen ble omrørt ved 0°C i 30 minutter, oppvarmet til romtemperatur og omrørt i 2 timer. Deretter, THF ble fjernet i vakuum. Residuet ble fortynnet med EtjO og ekstrahert med H2O (lx) og 0,5M NaOH (3x). De vandige ekstraktene ble surgjort med 2N HC1 og ekstrahert med CH2CI2 (3x). De samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (3,9 g). Flash kromatografi på silikagel under eluering med Heks:EtOAc (4:1) ga sub-tittelforbindelse (3,0 g, 61%) som en olje. 15.9 mmol; see step (iv) above) in anhydrous THF (400 mL) under a nitrogen atmosphere at less than 0°C (ice-MeOH bath) was added sodium ethoxide (4.5 g, 63.6 mmol). The cold solution was stirred for 40 minutes and ethyl formate (8.1 g, 111.3 mmol) was added. The solution was stirred at 0°C for 30 minutes, warmed to room temperature and stirred for 2 hours. Then, the THF was removed in vacuo. The residue was diluted with Et 2 O and extracted with H 2 O (1x) and 0.5 M NaOH (3x). The aqueous extracts were acidified with 2N HCl and extracted with CH 2 Cl 2 (3x). The combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound (3.9 g). Flash chromatography on silica gel eluting with Hex:EtOAc (4:1) gave the sub-title compound (3.0 g, 61%) as an oil.

<*>H NMR (300 MHz, CDCI3, blanding av isomerer): A 12,30 og 12,25 (s, 1H), 7,39 og 7,34 (s, 1H), 7,21 (s, 1H), 7,17 (s, 1H), 7,08 (s, 1H), 4,27-4,37 (q, 7 = 8 Hz, 2H), 1,28-1,38 <*>H NMR (300 MHz, CDCl 3 , mixture of isomers): A 12.30 and 12.25 (s, 1H), 7.39 and 7.34 (s, 1H), 7.21 (s, 1H ), 7.17 (s, 1H), 7.08 (s, 1H), 4.27-4.37 (q, 7 = 8 Hz, 2H), 1.28-1.38

(t,7=8Hz, 3H) (t,7=8Hz, 3H)

(vi) Ph( 3- Cl)( 5- OCFO-(/ ?. S) CH( CH, OH) C( Q) OEt (vi) Ph( 3- Cl)( 5- OCFO-(/ ?. S) CH( CH, OH) C( Q) OEt

Til en løsning av Ph(3-Cl)(5-OCF3)-(Æ,S)CH(CHO)C(0)OEt (3,0 g, To a solution of Ph(3-Cl)(5-OCF3)-(Æ,S)CH(CHO)C(0)OEt (3.0 g,

9,66 mmol; se trinn (v) ovenfor) i MeOH (200 ml) ved -10°C (is-MeOH bad) ble det tilsatt natrium-borhydrid (0,7 g, 19,32 mmol) porsjonsvis over 5 min. Løsningen ble omrørt ved -10°C i 45 minutter og ytterligere natrium-borhydrid (0,4 g) ble tilsatt. Etter ytterligere 15 minutter, ble reaksjonen stanset med vandig ammoniumklorid, gjort svakt sur med 2N HC1 og MeOH ble fjernet i vakuum. Residuet ble fortynnet med EtOAc og vasket med H2O (lx), vandig NaHC03 (lx) og saltvann (lx). De organiske faser ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse. Flash kromatografi på silikagel under eluering med Heks:EtOAc (5:1) ga sub-tittelforbindelse (2,0 g, 66%) som en olje. 9.66 mmol; see step (v) above) in MeOH (200 ml) at -10°C (ice-MeOH bath) sodium borohydride (0.7 g, 19.32 mmol) was added portionwise over 5 min. The solution was stirred at -10°C for 45 minutes and additional sodium borohydride (0.4 g) was added. After an additional 15 min, the reaction was quenched with aqueous ammonium chloride, slightly acidified with 2N HCl and the MeOH was removed in vacuo. The residue was diluted with EtOAc and washed with H 2 O (lx), aqueous NaHCO 3 (lx) and brine (lx). The organic phases were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give crude sub-title compound. Flash chromatography on silica gel eluting with Hex:EtOAc (5:1) gave the sub-title compound (2.0 g, 66%) as an oil.

'H NMR (300 MHz, CDCI3): A 7,26 (s, 1H), 7,19 (s, 1H), 7,07 (s, 1H), 4,16-4,28 (m, 2H), 4,04-4,15 (m, 1H), 3,76-3,94 (m, 2H), 2,33 (t, 7 = 6 Hz, 1H), 1,18-1,30 (t, 7 = 8 Hz, 3H) 1 H NMR (300 MHz, CDCl 3 ): Δ 7.26 (s, 1H), 7.19 (s, 1H), 7.07 (s, 1H), 4.16-4.28 (m, 2H) , 4.04-4.15 (m, 1H), 3.76-3.94 (m, 2H), 2.33 (t, 7 = 6 Hz, 1H), 1.18-1.30 (t , 7 = 8 Hz, 3H)

(vii) Ph( 3- Cl)( 5- OCF0-(/ g. 5) CH( CH2OH) C( O) OH (vii) Ph( 3- Cl)( 5- OCF0-(/ g. 5) CH( CH2OH) C( O) OH

Til en løsning av Ph(3-Cl)(5-OCF3)-(Æ,S)CH(CH2OH)C(0)OEt (2,0 g, To a solution of Ph(3-Cl)(5-OCF3)-(Æ,S)CH(CH2OH)C(0)OEt (2.0 g,

6,24 mmol; se trinn (vi) ovenfor) i THF (50 ml) og H20 (25 ml) ble tilsatt litiumhydroksydmonohydrat (0,5 g, 12,48 mmol). Løsningen ble omrørt ved romtemperatur i 1 time og THF ble fjernet 1 vakuum. Residuet ble fortynnet med H20, deretter vasket med CHCI3 (2x) og vaskevæskene kastet. Det basiske vandige lag ble surgjort med 2N HC1 og ekstrahert med CHC13 (4x). CHCI3 ekstrakter ble tørket (Na2SC«4), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (1,5 g) som en olje. Flash kromatografi på silikagel under eluering med CHCl3:MeOH:konsentrert NH4OH (gradient fra 7,0:2,5:0,5 til 6:3:1) ga ammoniumsalt av sub-tittelforbindelse (1,1 6.24 mmol; see step (vi) above) in THF (50 mL) and H 2 O (25 mL) was added lithium hydroxide monohydrate (0.5 g, 12.48 mmol). The solution was stirred at room temperature for 1 hour and the THF was removed under vacuum. The residue was diluted with H 2 O, then washed with CHCl 3 (2x) and the washings discarded. The basic aqueous layer was acidified with 2N HCl and extracted with CHCl 3 (4x). CHCl 3 extracts were dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give crude sub-title compound (1.5 g) as an oil. Flash chromatography on silica gel eluting with CHCl3:MeOH:conc NH4OH (gradient from 7.0:2.5:0.5 to 6:3:1) gave the ammonium salt of sub-title compound (1.1

g). Ammoniumsaltet ble fordelt mellom IN HC1 og CHCI3. De organiske faser ble tørket (Na2SC<4), filtrert og konsentrert 1 vakuum, hvilket gir sub-tittelforbindelsen (også kjent g). The ammonium salt was partitioned between IN HCl and CHCl 3 . The organic phases were dried (Na 2 SC<4), filtered and concentrated in vacuo to give the sub-title compound (also known

som 3-klor-5-trifluormetoksytropin syre) som en olje (1,1 g, 62%). as 3-chloro-5-trifluoromethoxytropinic acid) as an oil (1.1 g, 62%).

'H NMR (300 MHz, CD3OD):A 7,41 (s, 1H), 7,27 (s, 1H), 7,24 (s, 1H), 4,03 (m, 1H), 3,75-3,87 (m, 2H) 1 H NMR (300 MHz, CD 3 OD): Δ 7.41 (s, 1H), 7.27 (s, 1H), 7.24 (s, 1H), 4.03 (m, 1H), 3.75 -3.87 (m, 2H)

(viii) Ph( 3- Cl)( 5- OCF0-( 5) CH( CH7QH) C( O)- Aze- Pab( Teoc) ( a) og Ph( 3- Cn( 5- OCFO-(/ g) CH( CH7QH) C( Q)- Aze- Pab( Teoc)( b) (viii) Ph( 3- Cl)( 5- OCF0-( 5) CH( CH7QH) C( O)- Aze- Pab( Teoc) ( a) and Ph( 3- Cn( 5- OCFO-(/ g) CH( CH7QH) C( Q)- Aze- Pab( Teoc)( b)

Til en løsning av Ph(3-Cl)(5-OCF3)-(/?,S)CH(CH2OH)C(0)OH (0,65 g, To a solution of Ph(3-Cl)(5-OCF3)-(/?,S)CH(CH2OH)C(0)OH (0.65 g,

2,28 mmol; se trinn (vii) ovenfor) i DMF ved mindre enn 0°C (is-MeOH bad) ble det tilsatt H-Aze-Pab(Teoc) (0,90 g, 2,39 mmol), collidin (0,71 g, 5,70 mmol) og PyBOP (1,31 g, 2,51 mmol). Den resulterende løsning ble omrørt ved mindre enn 0°C i 1 time, oppvarmet til romtemperatur og omrørt i 1 time. DMF ble deretter fjernet i vakuum. Residuet ble fortynnet med EtOAc og vasket med fortynnet vandig HC1 (lx), saltvann (lx), vandig NaHC03 (lx) og saltvann (lx). De organiske faser ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir rå sub-tittelforbindelse (2,1 g) som en blanding av diastereomerene. Flash kromatografi (3x) på silikagel under eluering først med EtOAc:MeOH (95:5) deretter med CH2Cl2:MeOH (97:3) og deretter med CH2Cl2:MeOH 2.28 mmol; see step (vii) above) in DMF at less than 0 °C (ice-MeOH bath) was added H-Aze-Pab(Teoc) (0.90 g, 2.39 mmol), collidine (0.71 g , 5.70 mmol) and PyBOP (1.31 g, 2.51 mmol). The resulting solution was stirred at less than 0°C for 1 hour, warmed to room temperature and stirred for 1 hour. The DMF was then removed in vacuo. The residue was diluted with EtOAc and washed with dilute aqueous HCl (lx), brine (lx), aqueous NaHCO 3 (lx) and brine (lx). The organic phases were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the crude sub-title compound (2.1 g) as a mixture of the diastereomers. Flash chromatography (3x) on silica gel eluting first with EtOAc:MeOH (95:5) then with CH2Cl2:MeOH (97:3) and then with CH2Cl2:MeOH

(95:5) ga sub-tittelforbindelse diastereomer (a) (0,51 g, 35%) og diastereomer (b) (0,45 g, 31%) som knusbart skum. (95:5) gave sub-title compound diastereomer (a) (0.51 g, 35%) and diastereomer (b) (0.45 g, 31%) as crushable foam.

For sub-tittelforbindelse diastereomer (a) For sub-title compound diastereomer (a)

<!>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) A 7,79-7,85 (d, J = 8 Hz, 2H), 7,22-7,49 (m, 5H), 5,17-4,77 (m, 1H), 4,53-4,18 (m, 4H), 3,58-4,11 (m, 5H), 2,47-2,73 (m, 1H), 2,11-2,34 (m, 1H), 1,08-1,12 (m, 2H), 0,07 (s, 9H) <!>H NMR (300 MHz, CD3OD, complex mixture of rotamers) A 7.79-7.85 (d, J = 8 Hz, 2H), 7.22-7.49 (m, 5H), 5, 17-4.77 (m, 1H), 4.53-4.18 (m, 4H), 3.58-4.11 (m, 5H), 2.47-2.73 (m, 1H), 2.11-2.34 (m, 1H), 1.08-1.12 (m, 2H), 0.07 (s, 9H)

MS (m/z) 643 (M + 1)<+>MS (m/z) 643 (M + 1)<+>

(ix) Ph( 3- Cl)( 5- OCFj)-( 5) CH( CH2OH) C( 0)- Aze- Pab x TFA (ix) Ph( 3- Cl)( 5- OCFj)-( 5) CH( CH2OH) C( 0)- Aze- Pab x TFA

Ph(3-Cl)(5-OCF3)-(S)CH(CH2OH)C(0)-Aze-Pab(Teoc), (78 mg, 0,121 mmol; se trinn (viii) ovenfor - diastereomer (a)), ble oppløst i 5 ml trifluoreddiksyre. Etter 10 minutter, var reaksjonen over og løsningsmidlet ble avdampet. Residuet ble fryse tørket fra vann og acetonitril, hvilket gir det ønskede produkt. Utbytte: 70 mg (94%). Ph(3-Cl)(5-OCF3)-(S)CH(CH2OH)C(0)-Aze-Pab(Teoc), (78 mg, 0.121 mmol; see step (viii) above - diastereomer (a)) , was dissolved in 5 ml of trifluoroacetic acid. After 10 minutes, the reaction was over and the solvent was evaporated. The residue was freeze dried from water and acetonitrile to give the desired product. Yield: 70 mg (94%).

MS (m/z) 483 (M -1)"; 485 (M + 1)<+>MS (m/z) 483 (M -1)"; 485 (M + 1)<+>

<1>H-NMR(400 MHz; D20) rotamerer 1:1:8 8,83 (bt, 1H), 7,79 (d, 1H), 7,72 (d, 1H), 7,54 (d, 1H), 7,43 (d, 2H), 7,35 (m, 1H, rotamer), 7,28 (m, 1H, rotamer), 7,20 (m, 1H, rotamer), 7,05 (m, 1H, rotamer), 5,22 (m, 1H, rotamer), 4,83 (m, 1H, rotamer), 4,57 (m, 2H, rotamer), 4,38 (m, 2H, rotamer), 4,3-3,7 (m, 5H), 2,77 (m, 1H, rotamer), 2,55 (m, 1H, rotamer), 2,27 (m, 1H) <1>H-NMR(400 MHz; D20) rotamers 1:1:8 8.83 (bt, 1H), 7.79 (d, 1H), 7.72 (d, 1H), 7.54 (d , 1H), 7.43 (d, 2H), 7.35 (m, 1H, rotamer), 7.28 (m, 1H, rotamer), 7.20 (m, 1H, rotamer), 7.05 ( m, 1H, rotamer), 5.22 (m, 1H, rotamer), 4.83 (m, 1H, rotamer), 4.57 (m, 2H, rotamer), 4.38 (m, 2H, rotamer) , 4.3-3.7 (m, 5H), 2.77 (m, 1H, rotamer), 2.55 (m, 1H, rotamer), 2.27 (m, 1H)

<13>C-NMR (100 MHz; D20): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,9, 172,2,172,0,171,8,166,9 <13>C-NMR (100 MHz; D20): (carbonyl and/or amidine carbon atoms, rotamers) 8 172.9, 172.2,172.0,171.8,166.9

Eksempel 11 Example 11

Ph( 3- Cl)( 5- OCF0-( 5) CH( CH2OH) C( O)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCF0-( 5) CH( CH2OH) C( O)- Aze- Pab( OMe)

(i) Ph( 3- Cl)( 5- OCFO-( S) CH( CH, OH) C( 0)- Aze- Pab( OMe. Teoc) (i) Ph( 3- Cl)( 5- OCFO-( S) CH( CH, OH) C( 0)- Aze- Pab( OMe. Teoc)

Ph(3-Cl)(5-OCF3)-(S)CH(CH2OH)C(0)-Aze-Pab(Teoc) (100 mg, 0,155 mmol; se Eksempel 10(viii) ovenfor), ble oppløst i 12 ml tetrahydrofuran. O-metylhydroksylamin-hydroklorid (44 mg, Ph(3-Cl)(5-OCF3)-(S)CH(CH2OH)C(0)-Aze-Pab(Teoc) (100 mg, 0.155 mmol; see Example 10(viii) above), was dissolved in 12 ml of tetrahydrofuran. O-methylhydroxylamine hydrochloride (44 mg,

0,53 mmol), ble tilsatt og reaksjonsblandingen ble oppvarmet ved 50°C natten over. 0.53 mmol), was added and the reaction mixture was heated at 50°C overnight.

Reaksjonsblandingen ble inndampet og residuet renset ved preparativ HPLC (CH3CN/0,1 M NH4OAC (70/30)). Fraksjoner ble inndampet og residuet oppløst i en liten mengde av acetonitril og vann og fryse tørket. Fryse tørkingen ble gjentatt én gang. Utbytte: 80 mg (76%) av rent materiale. The reaction mixture was evaporated and the residue purified by preparative HPLC (CH 3 CN/0.1 M NH 4 OAC (70/30)). Fractions were evaporated and the residue dissolved in a small amount of acetonitrile and water and freeze dried. Freeze drying was repeated once. Yield: 80 mg (76%) of pure material.

'H-NMR(400 MHz; CD3OD) rotamerer: 8 7,5-7,4 (m, 3H), 7,35-7,2 (m, 4H), 5,15 (m, 1H, mindre rotamer), 4,74 (m, 1H, hoved rotamer), 4,5-4,25 (m, 3H), 4,2-3,95 (m, 4H), 3,91 (b, 3H), 3,9-3,6 (m, 2H), 2,63 (m, 1H, mindre rotamer), 2,50 (m, 1H, hoved rotamer), 2,3-2,1 (m, 1H), 0,95 (m, 2H), 0,02 (s, 9H, hoved rotamer), 0,01 (s, 9H, mindre rotamer) 'H-NMR(400 MHz; CD3OD) rotamer: δ 7.5-7.4 (m, 3H), 7.35-7.2 (m, 4H), 5.15 (m, 1H, minor rotamer) , 4.74 (m, 1H, main rotamer), 4.5-4.25 (m, 3H), 4.2-3.95 (m, 4H), 3.91 (b, 3H), 3, 9-3.6 (m, 2H), 2.63 (m, 1H, minor rotamer), 2.50 (m, 1H, major rotamer), 2.3-2.1 (m, 1H), 0, 95 (m, 2H), 0.02 (s, 9H, major rotamer), 0.01 (s, 9H, minor rotamer)

(ii) Ph( 3- Cn( 5- OCFj)-( 5) CH( CH9QH) C( 0)- Aze- Pab( OMe) (ii) Ph( 3- Cn( 5- OCFj)-( 5) CH( CH9QH) C( 0)- Aze- Pab( OMe)

Ph(3-Cl)(5-OCF3)-(S)CH(CH2OH)C(0)-Aze-Pab(OMe, Teoc), (80 mg, 0,12 mmol; se trinn (i) ovenfor), ble oppløst i 1 ml metylenklorid og avkjølt i et isbad. Trifluoreddiksyre, 3 ml, ble tilsatt og reaksjonskolben ble holdt i isbadet i to timer. Blandingen ble inndampet og oppløst i etylacetat og vasket tre ganger med NaHC03 (vandig) deretter med vann og saltvann. Den organiske fasen ble tørket (Na2SC«4), filtrert og inndampet. Residuet ble fryse tørket fra en liten mengde av acetonitril og vann. Utbytte: 60 mg (95%) av rent tittel produkt. Ph(3-Cl)(5-OCF3)-(S)CH(CH2OH)C(0)-Aze-Pab(OMe, Teoc), (80 mg, 0.12 mmol; see step (i) above), was dissolved in 1 ml of methylene chloride and cooled in an ice bath. Trifluoroacetic acid, 3 ml, was added and the reaction flask was kept in the ice bath for two hours. The mixture was evaporated and dissolved in ethyl acetate and washed three times with NaHCO 3 (aq) then with water and brine. The organic phase was dried (Na 2 SC 4 ), filtered and evaporated. The residue was freeze dried from a small amount of acetonitrile and water. Yield: 60 mg (95%) of pure title product.

MS (m/z) 528 (M -1)_; 531 (M + 1)<+>MS (m/z) 528 (M -1)_; 531 (M + 1)<+>

<1>H-NMR(500 MHz; CD3OD) rotamerer: 8 7,65-7,55 (m, 3H, rotamerer), 7,45 (m, 1H, hoved rotamer), 7,4-7,2 (m, 4H), 5,15 (m, 1H, mindre rotamer), 4,74 (m, 1H, hoved rotamer), 4,5-4,3 (m, 3H), 4,05-3,95 (m, 2H), 3,85 (m, 1H, hoved rotamer), 3,82 (s, 3H, hoved rotamer), 3,81 (s, 3H, mindre rotamer), 3,73 (m, 1H, hoved rotamer), 3,67 (m, 1H, mindre rotamer), 3,62 (m, 1H, mindre rotamer), 2,63 (m, 1H, mindre rotamer), 2,50 (m, 1H, hoved rotamer), 2,24 (m, 1H, hoved rotamer), 2,16 (m, 1H, mindre rotamer) <13>C-NMR (125 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 174,0,173,2,172,7,172,6,155,1 <1>H-NMR(500 MHz; CD3OD) rotamer: δ 7.65-7.55 (m, 3H, rotamer), 7.45 (m, 1H, main rotamer), 7.4-7.2 ( m, 4H), 5.15 (m, 1H, minor rotamer), 4.74 (m, 1H, major rotamer), 4.5-4.3 (m, 3H), 4.05-3.95 ( m, 2H), 3.85 (m, 1H, major rotamer), 3.82 (s, 3H, major rotamer), 3.81 (s, 3H, minor rotamer), 3.73 (m, 1H, major rotamer), 3.67 (m, 1H, minor rotamer), 3.62 (m, 1H, minor rotamer), 2.63 (m, 1H, minor rotamer), 2.50 (m, 1H, major rotamer) , 2.24 (m, 1H, major rotamer), 2.16 (m, 1H, minor rotamer) <13>C-NMR (125 MHz; CD3OD): (carbonyl and/or amidine carbons, rotamers) 174,0,173, 2,172,7,172,6,155,1

Eksempel 12 Example 12

Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab( OMe)

(i) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe. Teoc) (i) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe. Teoc)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,40 g, 0,65 mmol; se Eksempel l(ix) ovenfor), ble oppløst i 20 ml acetonitril og 0,50 g (6,0 mmol) O-metyl hydroksylamin-hydroklorid ble tilsatt. Blandingen ble oppvarmet ved 70°C i 2 timer. Løsningsmidlet ble avdampet og residuet ble fordelt mellom vann og etylacetat. Den vandige fasen ble ekstrahert to ganger mer med etylacetat og den samlede organiske fase ble vasket med vann, saltvann, tørket (Na2S04), filtrert og inndampet. Utbytte: 0,41 g (91%). Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.40 g, 0.65 mmol; see Example 1(ix) above) , was dissolved in 20 ml of acetonitrile and 0.50 g (6.0 mmol) of O-methyl hydroxylamine hydrochloride was added. The mixture was heated at 70°C for 2 hours. The solvent was evaporated and the residue was partitioned between water and ethyl acetate. The aqueous phase was extracted twice more with ethyl acetate and the combined organic phase was washed with water, brine, dried (Na 2 SO 4 ), filtered and evaporated. Yield: 0.41 g (91%).

'H-NMR (400 MHz; CDC13): 5 7,83 (bt, 1H), 7,57 (bs, 1H), 7,47 (d, 2H), 7,30 (d, 2H), 7,20 (m, 1H), 7,14 (m, 1H), 7,01 (m, 1H), 6,53 (t, 1H), 4,89 (s, 1H), 4,87 (m, 1H), 4,47 (m, 2H), 4,4-4,2 (b, 1H), 4,17-4,1 (m, 3H), 3,95 (s, 3H), 3,67 (m, 1H), 2,68 (m, 1H), 2,42 (m,lH) 0,97 (m, 2H), 0,01 (s, 9H). 1H-NMR (400 MHz; CDCl 3 ): δ 7.83 (bt, 1H), 7.57 (bs, 1H), 7.47 (d, 2H), 7.30 (d, 2H), 7, 20 (m, 1H), 7.14 (m, 1H), 7.01 (m, 1H), 6.53 (t, 1H), 4.89 (s, 1H), 4.87 (m, 1H ), 4.47 (m, 2H), 4.4-4.2 (b, 1H), 4.17-4.1 (m, 3H), 3.95 (s, 3H), 3.67 ( m, 1H), 2.68 (m, 1H), 2.42 (m, 1H) 0.97 (m, 2H), 0.01 (s, 9H).

(ii) Ph( 3- Cl)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- Pab( QMe) (ii) Ph( 3- Cl)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- Pab( QMe)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe, Teoc) (0,40 g, 0,62 mmol; se trinn (i) ovenfor), ble oppløst i 5 ml TFA og fikk reagere i 30 min. TFA ble inndampet og residuet ble fordelt mellom etylacetat og NaHCC«3 (vandig). Den vandige fasen ble ekstrahert to ganger mer med etylacetat og den samlede organiske fase ble vasket med vann, saltvann, tørket (Na2S04), filtrert og inndampet. Produktet ble fryse tørket fra vann/acetonitril. No rensning ble nødvendig.Utbytte: 0,28 g (85%). Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe, Teoc) (0.40 g, 0.62 mmol; see step (i) above ), was dissolved in 5 ml TFA and allowed to react for 30 min. The TFA was evaporated and the residue was partitioned between ethyl acetate and NaHCO3 (aq). The aqueous phase was extracted twice more with ethyl acetate and the combined organic phase was washed with water, brine, dried (Na 2 SO 4 ), filtered and evaporated. The product was freeze dried from water/acetonitrile. No purification was necessary. Yield: 0.28 g (85%).

'H-NMR (600 MHz; CDCI3): 6 7,89 (bt, 1H), 7,57 (d, 2H), 7,28 (d, 2H), 7,18 (m, 1H), 7,13 (m,lH), 6,99 (m, 1H), 6,51 (t, 1H), 4,88 (s, 1H), 4,87 (m, 1H), 4,80 (bs, 2H), 4,48 (dd, 1H), 4,43 (dd, 1H), 4,10 (m, 1H), 3,89 (s, 3H), 3,68 (m, 1H), 2,68 (m, 1H), 2,40 (m, 1H). <13>C-NMR (125 MHz; CDC13): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,9, 170,8,152,7,152,6 1H NMR (600 MHz; CDCl 3 ): δ 7.89 (bt, 1H), 7.57 (d, 2H), 7.28 (d, 2H), 7.18 (m, 1H), 7, 13 (m,lH), 6.99 (m, 1H), 6.51 (t, 1H), 4.88 (s, 1H), 4.87 (m, 1H), 4.80 (bs, 2H ), 4.48 (dd, 1H), 4.43 (dd, 1H), 4.10 (m, 1H), 3.89 (s, 3H), 3.68 (m, 1H), 2.68 (m, 1H), 2.40 (m, 1H). <13>C-NMR (125 MHz; CDC13): (carbonyl and/or amidine carbon atoms, rotamers) 8 172.9, 170.8,152.7,152.6

MS (m/z) 495 (M -1)\ 497 (M + 1)<+>MS (m/z) 495 (M -1)\ 497 (M + 1)<+>

Eksempel 13 Example 13

Ph( 3- OCHF9)-(/ ?) CH( OH) C( Q)- Aze- Pab x HOAc Ph( 3- OCHF9)-(/ ?) CH( OH) C( Q)- Aze- Pab x HOAc

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe) (13 mg, 0,026 mmol; se Eksempel 12 ovenfor) ble oppløst i abs. etanol (5 ml) og 30 mg 10% Pd/C ble tilsatt. Til slutt ble eddiksyre (5 (iL) tilsatt og blandingen ble hydrogenert ved atmosfærisk trykk i 20 timer. Blandingen ble filtrert gjennom Celite®, inndampet og renset ved reversert fase HPLC (0,1 M vandig ammoniumacetat/MeCN). De passende fraksjoner ble frysetørket, hvilket gir tittelforbindelsen som et hvitt, fast stoff: 8,5 mg (66%). Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe) (13 mg, 0.026 mmol; see Example 12 above) was dissolved in abs. ethanol (5 mL) and 30 mg of 10% Pd/C were added. Finally, acetic acid (5 (iL)) was added and the mixture was hydrogenated at atmospheric pressure for 20 h. The mixture was filtered through Celite®, evaporated and purified by reversed phase HPLC (0.1 M aqueous ammonium acetate/MeCN). The appropriate fractions were lyophilized to give the title compound as a white solid: 8.5 mg (66%).

<1>H-NMR(400 MHz; CD3OD) rotamerer: 8 7,73-7,78 (m, 2H), 7,55 (d, 2H), 7,19-7,43 (m, 3H), 7,06-7,13 (m, 1H), 6,83 (t, 1H, JHF = 74Hz, hoved rotamer), 6,81 (t, 1H, hoved rotamer), 5,20 (s, 1H, hoved rotamer), 5,19 (m, 1H, mindre rotamer), 5,15 (s, 1H, mindre rotamer), 4,78 (m, 1H, hoved rotamer), 4,4-4,6 (mange topper, 2H), 4,35 (m, 1H, hoved rotamer), 4,08 (m, 1H), 3,99 (m, 1H, mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,52 (m, 1H, hoved rotamer), 2,30 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer), 1,89 (s, 3H). 1 ^C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 173,7, 172,9,168,3. <1>H-NMR(400 MHz; CD3OD) rotamers: δ 7.73-7.78 (m, 2H), 7.55 (d, 2H), 7.19-7.43 (m, 3H), 7.06-7.13 (m, 1H), 6.83 (t, 1H, JHF = 74Hz, main rotamer), 6.81 (t, 1H, main rotamer), 5.20 (s, 1H, main rotamer), 5.19 (m, 1H, minor rotamer), 5.15 (s, 1H, minor rotamer), 4.78 (m, 1H, major rotamer), 4.4-4.6 (many peaks, 2H), 4.35 (m, 1H, major rotamer), 4.08 (m, 1H), 3.99 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2, 52 (m, 1H, major rotamer), 2.30 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer), 1.89 (s, 3H). 1 3 C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms, rotamers) δ 173.7, 172.9, 168.3.

MS (m/z) 433 (M+l)+ ; 431 (M-l)- MS (m/z) 433 (M+1) + ; 431 (M-l)-

Eksempel 14 Example 14

Ph( 3- OCFO-(/ g) CH( OH) C( Q)- Aze- Pab x TFA Ph( 3- OCFO-(/ g) CH( OH) C( Q)- Aze- Pab x TFA

Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab x TFA (34 mg, 0,057 mmol, fra Eksempel 6) ble oppløst i 5 ml etanol og 20 mg 10% Pd/C ble tilsatt. Blandingen ble hydrogenen ved atmosfærisk trykk natten over. Blandingen ble filtrert gjennom Celite®, inndampet og fryse tørket fra vann/acetonitril. Ph(3-Cl)(5-OCF3)-(/?)CH(OH)C(0)-Aze-Pab x TFA (34 mg, 0.057 mmol, from Example 6) was dissolved in 5 ml ethanol and 20 mg 10% Pd/C was added. The mixture was hydrogenated at atmospheric pressure overnight. The mixture was filtered through Celite®, evaporated and freeze dried from water/acetonitrile.

'H-NMR(400 MHz; CD3OD) rotamerer: 8 7,8-7,7 (m, 2H), 7,55 (m, 2H), 7,5-7,2 (m, 4H), 5,24 (s, 1H, hoved rotamer), 5,23 (m, 1H, mindre rotamer), 5,18 (s, 1H, mindre rotamer), 4,77 (m, 1H, hoved rotamer), 4,6-4,45 (m, 2H), 4,36 (m, 1H, hoved rotamer), 4,08 (m, 1H), 3,99 (m, 1H, mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,52 (m, 1H, hoved rotamer), 2,30 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer). 1H-NMR(400 MHz; CD3OD) rotamers: δ 7.8-7.7 (m, 2H), 7.55 (m, 2H), 7.5-7.2 (m, 4H), 5, 24 (s, 1H, major rotamer), 5.23 (m, 1H, minor rotamer), 5.18 (s, 1H, minor rotamer), 4.77 (m, 1H, major rotamer), 4.6- 4.45 (m, 2H), 4.36 (m, 1H, major rotamer), 4.08 (m, 1H), 3.99 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.52 (m, 1H, major rotamer), 2.30 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer).

<11>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 174,1,173,9,173,5,172,9,168,2. <11>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 174,1,173,9,173,5,172,9,168,2.

<19->F NMR (282 MHz; CD3OD): -59,8 og -59,9 (3F, henholdsvis mindre og hoved rotamer), <19->F NMR (282 MHz; CD3OD): -59.8 and -59.9 (3F, minor and major rotamer, respectively),

-77,4 (3F) indikerer at saltet er TFA. -77.4 (3F) indicates that the salt is TFA.

MS (m/z) 451,3 (M+l)<+>MS (m/z) 451.3 (M+1)<+>

Eksempel 15 Example 15

Ph( 3- Cl¥5- OCH?CF0-(/ g) CH( OH) C( O)- Aze- Pab x TFA Ph( 3- Cl¥5- OCH?CF0-(/ g) CH( OH) C( O)- Aze- Pab x TFA

(i) 3- klor- 5- trifluoretoksvbenzaldehyd (i) 3-chloro-5-trifluoroethoxybenzaldehyde

Til en magnetisk omrørt løsning av 3-klor-5-hydroksybenzaldehyd (2,0 g, 12,8 mmol; se Eksempel l(ii) ovenfor) og kaliumkarbonat (2,3 g, 16,6 mmol) i DMF (35 ml) under nitrogen ble tilsatt 2,2,2-trifluoretyl p-toluensulfonat (4,2 g, 16,6 mmol) ved romtemperatur. Blandingen ble oppvarmet til 110°C i 7 timer og deretter omrørt natten over ved romtemperatur. Reaksjonsblandingen ble avkjølt til 0°C, hellet i is-kald 2 N HC1 (100 ml) og ekstrahert med EtOAc (2 x 75 ml). De samlede organiske ekstrakter ble vasket med 0,5 N HC1 (2 x 50 ml), tørket (Na2S04), filtrert og konsentrert i vakuum. Den brune oljen ble kromatografert på silikagel under eluering med Heks:EtOAc (6:1), hvilket gir sub-tittelforbindelse (1,9 g, 61%) som en gul olje. To a magnetically stirred solution of 3-chloro-5-hydroxybenzaldehyde (2.0 g, 12.8 mmol; see Example 1(ii) above) and potassium carbonate (2.3 g, 16.6 mmol) in DMF (35 mL ) under nitrogen was added 2,2,2-trifluoroethyl p-toluenesulfonate (4.2 g, 16.6 mmol) at room temperature. The mixture was heated to 110°C for 7 hours and then stirred overnight at room temperature. The reaction mixture was cooled to 0°C, poured into ice-cold 2N HCl (100 mL) and extracted with EtOAc (2 x 75 mL). The combined organic extracts were washed with 0.5 N HCl (2 x 50 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with Hex:EtOAc (6:1) to give the sub-title compound (1.9 g, 61%) as a yellow oil.

'H NMR (300 MHz, CDC13) A 9,44 (s, 1H), 7,56 (s, 1H), 7,33 (s, 1H), 7,28 (s, 1H), 4,42 1 H NMR (300 MHz, CDCl 3 ) Δ 9.44 (s, 1H), 7.56 (s, 1H), 7.33 (s, 1H), 7.28 (s, 1H), 4.42

(q,7=8Hz, 2H) (q,7=8Hz, 2H)

(ii) Ph( 3- Cn( 5- OCH?CFO-( j?. 5) CH( OTMS) CN (ii) Ph( 3- Cn( 5- OCH?CFO-( j?. 5) CH( OTMS) CN

Til en løsning av 3-klor-5-trifluoretoksybenzaldehyd (5,2 g, 21,7 mmol; se trinn (i) ovenfor) og sinkjodid (1,7 g, 5,4 mmol) i CH2CI2 (200 ml) under nitrogen ble tilsatt trimetylsilylcyanid (4,3 g, 43,3 mmol) dråpevis via sprøyte ved 0°C. Blandingen ble omrørt ved 0°C i 3 timer og deretter fortynnet med H2O (150 ml). Det organiske laget ble separert, tørket (Na2S04), filtrert og konsentrert 1 vakuum, hvilket gir sub-tittelforbindelsen (6,9 g, 95%) som en gul olje som ble anvendt uten ytterligere rensning. To a solution of 3-chloro-5-trifluoroethoxybenzaldehyde (5.2 g, 21.7 mmol; see step (i) above) and zinc iodide (1.7 g, 5.4 mmol) in CH 2 Cl 2 (200 mL) under nitrogen Trimethylsilyl cyanide (4.3 g, 43.3 mmol) was added dropwise via syringe at 0°C. The mixture was stirred at 0°C for 3 h and then diluted with H 2 O (150 mL). The organic layer was separated, dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (6.9 g, 95%) as a yellow oil which was used without further purification.

'H NMR (300 MHz, CDCI3) A 7,27 (s, 1H), 6,98 (s, 2H), 5,44 (s, 1H), 4,38 (q, / = 8 Hz, 2H), 0,30 (s, 9 H) 1 H NMR (300 MHz, CDCl 3 ) Δ 7.27 (s, 1H), 6.98 (s, 2H), 5.44 (s, 1H), 4.38 (q, / = 8 Hz, 2H) , 0.30 (s, 9H)

(iii) Ph( 3- Cl¥5- OCH7CFj)-(/ ?. S) CH( OH) C( Q) OH (iii) Ph( 3- Cl¥5- OCH7CFj)-(/ ?. S) CH( OH) C( Q) OH

Konsentrert saltsyre (170 ml) ble satt til Ph(3-Cl)(5-OCH2CF3)-(/?,5)CH(OTMS)CN (6,9 g, 20,4 mmol; se trinn (ii) ovenfor) og omrørt ved 100°C i 1 time. Etter avkjøling til romtemperatur, ble reaksjonen ytterligere avkjølt til 0°C og gjort basisk langsomt med 3 N NaOH (300 ml). Denne blandingen ble vasket med Et20 (2 x 100 ml) og det vandige laget ble surgjort med 2 N HC1 (50 ml). Det vandige laget ble deretter ekstrahert med EtOAc (2 x 100 ml), tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelsen (5,3 g, 92%) som en blekgul olje som ble anvendt uten ytterligere rensning. Concentrated hydrochloric acid (170 mL) was added to Ph(3-Cl)(5-OCH2CF3)-(/?,5)CH(OTMS)CN (6.9 g, 20.4 mmol; see step (ii) above) and stirred at 100°C for 1 hour. After cooling to room temperature, the reaction was further cooled to 0°C and basified slowly with 3N NaOH (300 mL). This mixture was washed with Et 2 O (2 x 100 mL) and the aqueous layer was acidified with 2N HCl (50 mL). The aqueous layer was then extracted with EtOAc (2 x 100 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (5.3 g, 92%) as a pale yellow oil which was used without further purification .

'H NMR (300 MHz, CD3OD) A 7,18 (s, 1H), 7,07 (s, 1H), 7,02 (s, 1H), 5,13 (s, 1H), 4,58 1 H NMR (300 MHz, CD 3 OD) Δ 7.18 (s, 1H), 7.07 (s, 1H), 7.02 (s, 1H), 5.13 (s, 1H), 4.58

(q,7=8Hz, 2H) (q,7=8Hz, 2H)

(iv) Ph( 3- Cl¥5- OCH7CF0-(/ g) CH( OH) C( O) OH ( a) oe Ph( 3- Cl¥5- OCH?CFO-( 5) CH( OAc) C( Q) OH ( b) (iv) Ph( 3- Cl¥5- OCH7CF0-(/ g) CH( OH) C( O) OH ( a) oe Ph( 3- Cl¥5- OCH?CFO-( 5) CH( OAc) C (Q)OH (b)

En løsning av Ph(3-Cl)(5-OCH2CF3)-(Æ,S)CH(OH)C(0)OH (7,06 g, 24,8 mmol; se trinn (iii) ovenfor) og Lipase PS "Amano" (4,30 g) i vinylacetat (250 ml) og MTBE (250 ml) ble oppvarmet ved 70°C under nitrogen i 40 timer. Reaksjonsblandingen ble avkjølt til romtemperatur, enzymet ble fjernet ved filtrering vasking med EtOAc og filtratet konsentrert i vakuum. Kromatografi på silikagel under eluering med CHCl3:MeOH:Et3N (92:6:2) ga trietylaminsaltet av sub-tittelforbindelse (a) (3,02 g) som en gul olje. Saltet av sub-tittelforbindelse (a) ble oppløst i H20 (150 ml), surgjort med 2 N HC1 og ekstrahert med EtOAc (2 x 75 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelse (a) (2,18 g) som et gråhvitt, fast stoff. I tillegg, trietylaminsaltet av sub-tittelforbindelse (b) (4,73 g) ble oppnådd fra kolonnenkromatografi nevnt ovenfor. A solution of Ph(3-Cl)(5-OCH2CF3)-(Æ,S)CH(OH)C(0)OH (7.06 g, 24.8 mmol; see step (iii) above) and Lipase PS "Amano" (4.30 g) in vinyl acetate (250 mL) and MTBE (250 mL) was heated at 70°C under nitrogen for 40 hours. The reaction mixture was cooled to room temperature, the enzyme was removed by filtration, washing with EtOAc and the filtrate concentrated in vacuo. Chromatography on silica gel eluting with CHCl 3 :MeOH:Et 3 N (92:6:2) gave the triethylamine salt of sub-title compound (a) (3.02 g) as a yellow oil. The salt of sub-title compound (a) was dissolved in H 2 O (150 mL), acidified with 2 N HCl and extracted with EtOAc (2 x 75 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give sub-title compound (a) (2.18 g) as an off-white solid. In addition, the triethylamine salt of sub-title compound (b) (4.73 g) was obtained from the column chromatography mentioned above.

Data for sub-tittelforbindelse (a): Data for sub-title connection (a):

Sm.p.: 98-103°C Melting point: 98-103°C

<!>H NMR (300 MHz, CD3OD) A 7,18 (s, 1H), 7,07 (s, 1H), 7,02 (s, 1H), 5,13 (s, 1H), 4,58 (q,<y>=8Hz,2H). <!>H NMR (300 MHz, CD3OD) Δ 7.18 (s, 1H), 7.07 (s, 1H), 7.02 (s, 1H), 5.13 (s, 1H), 4, 58 (q,<y>=8Hz,2H).

<13>C NMR (75 MHz, CD3OD) A 175,4,159,6,144,6, 136,2,125,0 (q, J = 277 Hz), 121,8, 115,9,113,1,73,3, 67,0 (q, J =35 Hz) <13>C NMR (75 MHz, CD3OD) A 175.4,159.6,144.6, 136.2,125.0 (q, J = 277 Hz), 121.8, 115.9,113.1,73.3, 67, 0 (q, J = 35 Hz)

HPLC Analyse: 98,6%, >99% ee, Chiralcel OD Kolonne (97:3:0,5 Heks:EtOH:TFA mobil fase) HPLC Analysis: 98.6%, >99% ee, Chiralcel OD Column (97:3:0.5 Hex:EtOH:TFA mobile phase)

[I]<25>D = -81,5° (c = 1,0, MeOH) [I]<25>D = -81.5° (c = 1.0, MeOH)

APCI-MS: (M - 1) = 283 m/z APCI-MS: (M - 1) = 283 m/z

(v) Ph( 3- Cl)( 5- OCH7CFO-(/ tfCH( OH) C( 0)- Aze- Pab( Teoc) (v) Ph( 3- Cl)( 5- OCH7CFO-(/ tfCH( OH) C( 0)- Aze- Pab( Teoc)

Til en løsning av Ph(3-Cl)(5-OCH2CF3)-(Æ)CH(OH)C(0)OH (0,50 g, 1,8 mmol; se trinn (iv) ovenfor (forbindelse (a))) i DMF (20 ml) under nitrogen ble tilsatt H-Aze-Pab(Teoc) x HC1 (1,03 g, 2,3 mmol), PyBOP (1,01 g, 1,9 mmol) og DIPEA (0,57 g, 4,4 mmol) ved 0°C. Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur i 20 timer. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (10:1) og deretter med EtOAc:EtOH (10:1), hvilket gir sub-tittelforbindelsen (0,55 g, 48%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH2CF3)-(Æ)CH(OH)C(0)OH (0.50 g, 1.8 mmol; see step (iv) above (compound (a) )) in DMF (20 mL) under nitrogen was added H-Aze-Pab(Teoc) x HCl (1.03 g, 2.3 mmol), PyBOP (1.01 g, 1.9 mmol) and DIPEA (0 .57 g, 4.4 mmol) at 0°C. The reaction mixture was stirred at 0°C for 2 hours and then at room temperature for 20 hours. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl3:EtOH (10:1) and then with EtOAc:EtOH (10:1) to give the sub-title compound (0.55 g, 48% ) as a crushable white foam.

Sm.p.: 90-95°C Melting point: 90-95°C

Rf = 0,42 (10:1 CHCl3:EtOH) Rf = 0.42 (10:1 CHCl3:EtOH)

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) A 7,78-7,81 (m, 2H), 7,38-7,41 (m, 2H), 7,12-7,16 (m, 1H), 7,00-7,06 (m, 2H), 5,09-5,22 og 4,75-4,79 (m, 2H), 3,94-4,61 (m, 8H), 2,09-2,75 (m, 2H), 1,04-1,11 (m, 2H), 0,70 (s, 9H) 1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) Δ 7.78-7.81 (m, 2H), 7.38-7.41 (m, 2H), 7.12-7.16 (m , 1H), 7.00-7.06 (m, 2H), 5.09-5.22 and 4.75-4.79 (m, 2H), 3.94-4.61 (m, 8H) , 2.09-2.75 (m, 2H), 1.04-1.11 (m, 2H), 0.70 (s, 9H)

APCI-MS: (M + l) = 643m/z APCI-MS: (M + 1) = 643m/z

(vi) Ph( 3- ClX5- OCH9CF0-(/ g) CH( OH) C( O)- Aze- Pab x TFA (vi) Ph( 3- ClX5- OCH9CF0-(/ g) CH( OH) C( O)- Aze- Pab x TFA

Ph(3-Cl)(5-OCH2CF3)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,066 g, 0,103 mmol; se trinn (v) ovenfor), ble oppløst i 3 ml TFA og fikk reagere i 30 min. TFA ble inndampet og residuet ble fryse tørket fra vann/acetonitril, hvilket gir 0,060 g (94%) av tittelforbindelsen som dens TFA salt. Ph(3-Cl)(5-OCH2CF3)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.066 g, 0.103 mmol; see step (v) above), was dissolved in 3 ml TFA and allowed to react for 30 min. The TFA was evaporated and the residue was freeze dried from water/acetonitrile to give 0.060 g (94%) of the title compound as its TFA salt.

'H-NMR (400 MHz; CD3OD) rotamerer: 5 7,8-7,7 (m, 2H), 7,6-7,5 (m, 2H), 7,2-7,0 (m, 3H), 5,21 (m, 1H, mindre rotamer), 5,17 (s, 1H, hoved rotamer), 5,11 (s, 1H, mindre 1H-NMR (400 MHz; CD 3 OD) rotamers: δ 7.8-7.7 (m, 2H), 7.6-7.5 (m, 2H), 7.2-7.0 (m, 3H ), 5.21 (m, 1H, minor rotamer), 5.17 (s, 1H, major rotamer), 5.11 (s, 1H, minor

rotamer), 4,81 (m, 1H, hoved rotamer), 4,6-4,4 (m, 4H), 4,37 (m, 1H, hoved rotamer), 4,16 (m, 1H, hoved rotamer), 4,06 (m, 1H, mindre rotamer), 3,99 (m, 1H, mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,54 (m, 1H, hoved rotamer), 2,29 (m, 1H, hoved rotamer), 2,15 rotamer), 4.81 (m, 1H, main rotamer), 4.6-4.4 (m, 4H), 4.37 (m, 1H, main rotamer), 4.16 (m, 1H, main rotamer ), 4.06 (m, 1H, minor rotamer), 3.99 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.54 (m, 1H, major rotamer), 2.29 (m, 1H, main rotamer), 2.15

(m, 1H, mindre rotamer) (m, 1H, minor rotamer)

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 5 172,2,171,8,171,7,167,0. <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 5 172,2,171,8,171,7,167,0.

MS (m/z) 499,3 (M+l)<+>MS (m/z) 499.3 (M+1)<+>

Eksempel 16 Example 16

Ph( 3- Cl)( 5- OCH7CF0-(/ ?) CH( OH) C( O)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCH7CF0-(/ ?) CH( OH) C( O)- Aze- Pab( OMe)

Til en løsning av Ph(3-Cl)(5-OCH2CF3)-(/?)CH(OH)C(0)OH (0,48 g, 1,7 mmol; se Eksempel 15(iv) ovenfor (forbindelse (a)) i DMF (20 ml) under nitrogen ble tilsatt H-AzePab(OMe) x 2HC1 (0,74 g, 2,2 mmol), PyBOP (0,97 g, 1,9 mmol) og DIPEA (0,55 g, 4,2 mmol) ved 0°C. Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur i 20 timer. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (10:1) og deretter med EtOAc:EtOH (10:1), hvilket gir tittelforbindelsen (0,62 g, 69%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH2CF3)-(/?)CH(OH)C(0)OH (0.48 g, 1.7 mmol; see Example 15(iv) above (compound ( a)) in DMF (20 mL) under nitrogen was added H-AzePab(OMe) x 2HCl (0.74 g, 2.2 mmol), PyBOP (0.97 g, 1.9 mmol) and DIPEA (0, 55 g, 4.2 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 2 h and then at room temperature for 20 h. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl 3 : EtOH (10:1) and then with EtOAc:EtOH (10:1) to give the title compound (0.62 g, 69%) as a breakable white foam.

Sm.p.: 75-80°C Melting point: 75-80°C

Rf = 0,43 (10:1 CHCl3:EtOH) Rf = 0.43 (10:1 CHCl3:EtOH)

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) A 7,57-7,60 (m, 2H), 7,32-7,36 (m, 2H), 7,13-7,17 (m, 1H), 7,00-7,06 (m, 2H), 5,09-5,19 og 4,74-4,80 (m, 2H), 3,93-4,62 (m, 6H), 3,81 (s, 3H), 2,10-2,73 (m, 2H) 1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) Δ 7.57-7.60 (m, 2H), 7.32-7.36 (m, 2H), 7.13-7.17 (m , 1H), 7.00-7.06 (m, 2H), 5.09-5.19 and 4.74-4.80 (m, 2H), 3.93-4.62 (m, 6H) , 3.81 (s, 3H), 2.10-2.73 (m, 2H)

APCI-MS: (M+l) = 529m/z APCI-MS: (M+1) = 529m/z

Eksempel 17 Example 17

Ph( 3- Cl)( 5- OCH7CHF7)-(/ ?) CH( OH) C( Q)- Aze- Pab x TFA Ph( 3- Cl)( 5- OCH7CHF7)-(/ ?) CH( OH) C( Q)- Aze- Pab x TFA

(i) 2. 2- difluoretvlester metansulfonsvre (i) 2. 2- difluoroethyl ester methanesulfonic acid

Til en magnetisk omrørt løsning av 2,2-difluoretanol (1,52 g, 18,5 mmol) i CH2CI2 (20 ml) under nitrogen ble det tilsatt trietylamin (5,61 g, 55,5 mmol) og metansulfonylklorid (2,54 g, 22,2 mmol) ved 0°C. Blandingen ble omrørt ved 0°C i 1,5 timer, fortynnet med CH2CI2 (50 ml) og vasket med 2 N HC1 (50 ml). Det vandige laget ble ekstrahert med CH2C12 (30 ml) og de samlede organiske ekstrakter vasket med saltvann (30 ml), tørket (Na2SC<4), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelsen (2,52 g, 85%) som en gul olje som ble anvendt uten ytterligere rensning. To a magnetically stirred solution of 2,2-difluoroethanol (1.52 g, 18.5 mmol) in CH 2 Cl 2 (20 mL) under nitrogen was added triethylamine (5.61 g, 55.5 mmol) and methanesulfonyl chloride (2, 54 g, 22.2 mmol) at 0°C. The mixture was stirred at 0°C for 1.5 h, diluted with CH 2 Cl 2 (50 mL) and washed with 2N HCl (50 mL). The aqueous layer was extracted with CH 2 Cl 2 (30 mL) and the combined organic extracts washed with brine (30 mL), dried (Na 2 SC<4), filtered and concentrated in vacuo to give the sub-title compound (2.52 g, 85% ) as a yellow oil which was used without further purification.

'H NMR (300 MHz, CDC13) A 6,02 (tt, J = 3 Hz, J = 55 Hz, 1H), 4,39 (dt, J = 3 Hz, / = 13 Hz, 2H), 3,13 (s, 3H) 1 H NMR (300 MHz, CDCl 3 ) Δ 6.02 (tt, J = 3 Hz, J = 55 Hz, 1H), 4.39 (dt, J = 3 Hz, / = 13 Hz, 2H), 3, 13 (p, 3H)

(ii) 3 - klor- 5- difluoretoks vbenzaldehvd (ii) 3 - chloro- 5-difluoroethoxy vbenzaldehvd

Til en løsning av 3-klor-5-hydroksybenzaldehyd (1,50 g, 9,6 mmol; se Eksempel l(ii) ovenfor) og kaliumkarbonat (1,72 g, 12,5 mmol) i DMF (10 ml) under nitrogen ble tilsatt en løsning av 2,2-difluoretylester metansulfonsyre (2,0 g, 12,5 mmol; se trinn (i) ovenfor) i DMF (10 ml) dråpevis ved romtemperatur. Blandingen ble oppvarmet til 100°C i 6 timer og deretter omrørt natten over ved romtemperatur. Reaksjonsblandingen ble avkjølt til 0°C, hellet i is-kald 2 N HC1 (100 ml) og ekstrahert med EtOAc (2 x 75 ml). De samlede organiske ekstrakter ble vasket med 0,5 N HC1 (2 x 50 ml), tørket (Na2S04), filtrert og konsentrert i vakuum. Den brune oljen ble kromatografert på silikagel under eluering med Heks:EtOAc (5:1), hvilket gir sub-tittelforbindelsen (1,35 g, 64%) som en gul olje. To a solution of 3-chloro-5-hydroxybenzaldehyde (1.50 g, 9.6 mmol; see Example 1(ii) above) and potassium carbonate (1.72 g, 12.5 mmol) in DMF (10 mL) under nitrogen was added to a solution of 2,2-difluoroethyl ester methanesulfonic acid (2.0 g, 12.5 mmol; see step (i) above) in DMF (10 mL) dropwise at room temperature. The mixture was heated to 100°C for 6 hours and then stirred overnight at room temperature. The reaction mixture was cooled to 0°C, poured into ice-cold 2N HCl (100 mL) and extracted with EtOAc (2 x 75 mL). The combined organic extracts were washed with 0.5 N HCl (2 x 50 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with Hex:EtOAc (5:1) to give the sub-title compound (1.35 g, 64%) as a yellow oil.

'H NMR (300 MHz, CDC13) A 9,92 (s, 1H), 7,52 (s, 1H), 7,31 (s, 1H), 7,22 (s, 1H), 6,12 (tt, J = 3 Hz, J = 55 Hz, 1H), 4,26 (dt, J = 3 Hz, J = 15 Hz, 2H) 1 H NMR (300 MHz, CDCl 3 ) Δ 9.92 (s, 1H), 7.52 (s, 1H), 7.31 (s, 1H), 7.22 (s, 1H), 6.12 ( tt, J = 3 Hz, J = 55 Hz, 1H), 4.26 (dt, J = 3 Hz, J = 15 Hz, 2H)

(iii) Ph( 3- Cl)( 5- OCHzCHFz)-(/ g. 5) CH( OTMS) CN (iii) Ph( 3- Cl)( 5- OCHzCHFz)-(/ g. 5) CH( OTMS) CN

Til en løsning av 3-klor-5-difluoretoksybenzaldehyd (1,35 g, 6,1 mmol; se trinn (ii) ovenfor) og sinkjodid (0,48 g, 1,5 mmol) i CH2CI2 (50 ml) ble det tilsatt trimetylsilylcyanid (1,21 g, 12,2 mmol) dråpevis ved 0°C under nitrogen. Blandingen ble omrørt ved 0°C i 3 timer, deretter fortynnet med H20 (50 ml). Det organiske laget ble separert, tørket (Na2S04), filtrert og konsentrert 1 vakuum, hvilket gir sub-tittelforbindelse (1,85 g, 95%) som en brun olje som ble anvendt uten ytterligere rensning. To a solution of 3-chloro-5-difluoroethoxybenzaldehyde (1.35 g, 6.1 mmol; see step (ii) above) and zinc iodide (0.48 g, 1.5 mmol) in CH 2 Cl 2 (50 mL) was added added trimethylsilyl cyanide (1.21 g, 12.2 mmol) dropwise at 0°C under nitrogen. The mixture was stirred at 0°C for 3 h, then diluted with H 2 O (50 mL). The organic layer was separated, dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (1.85 g, 95%) as a brown oil which was used without further purification.

'H NMR (300 MHz, CDC13) A 7,13 (s, 1H), 6,94 (s, 2H), 6,10 (tt, / = 3 Hz, J = 55 Hz, 1H), 5,43 (s, 1H), 4,20 (dt, J = 3 Hz, J = 15 Hz, 2H), 0,28 (s, 9H) 1 H NMR (300 MHz, CDCl 3 ) Δ 7.13 (s, 1H), 6.94 (s, 2H), 6.10 (tt, / = 3 Hz, J = 55 Hz, 1H), 5.43 (s, 1H), 4.20 (dt, J = 3 Hz, J = 15 Hz, 2H), 0.28 (s, 9H)

(iv) Ph( 3- Cl)( 5- OCH, CHF,)-(/ g. S) CH( OH) aO) OH (iv) Ph( 3- Cl)( 5- OCH, CHF,)-(/ g. S) CH( OH) aO) OH

Konsentrert saltsyre (60 ml) ble satt til Ph(3-Cl)(5-OCH2CHF2)-(Æ,S)CH(OTMS)CN (1,85 g, 5,8 mmol; se trinn (iii) ovenfor) og omrørt ved 100°C i 1 time. Etter avkjøling til romtemperatur, ble reaksjonen ytterligere avkjølt til 0°C, gjort basisk langsomt med 3 N NaOH (-180 ml) og vasket med Et20 (2 x 75 ml). Det vandige laget ble surgjort med 2 N HC1 (20 ml) og ekstrahert med EtOAc (2 x 75 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert / vakuum, hvilket gir sub-tittelforbindelsen (1,50 g, 97%) som et blekgult, fast stoff som ble anvendt uten ytterligere rensning. Concentrated hydrochloric acid (60 mL) was added to Ph(3-Cl)(5-OCH2CHF2)-(Æ,S)CH(OTMS)CN (1.85 g, 5.8 mmol; see step (iii) above) and stirred at 100°C for 1 hour. After cooling to room temperature, the reaction was further cooled to 0°C, basified slowly with 3 N NaOH (-180 mL) and washed with Et 2 O (2 x 75 mL). The aqueous layer was acidified with 2N HCl (20 mL) and extracted with EtOAc (2 x 75 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated/vacuum to give the sub-title compound (1.50 g, 97%) as a pale yellow solid which was used without further purification.

<*>H NMR (300 MHz, CD3OD) A 7,15 (s, 1H), 7,05 (s, 1H), 6,98 (s, 1H), 6,19 (tt, 7 = 4 Hz, J = 55 Hz, 1H), 5,12 (s, 1H), 4,25 (dt, J = 4 Hz, J = 17 Hz, 2H) <*>H NMR (300 MHz, CD3OD) Δ 7.15 (s, 1H), 7.05 (s, 1H), 6.98 (s, 1H), 6.19 (tt, 7 = 4 Hz, J = 55 Hz, 1H), 5.12 (s, 1H), 4.25 (dt, J = 4 Hz, J = 17 Hz, 2H)

(v) Ph( 3- Cl)( 5- OCHzCHF?>( S) CH( OAc) C( 0) OH ( a) og Ph( 3- Cn( 5- OCH7CHF9)-(/ g) CH( OH) C( Q) OH ( b) (v) Ph( 3- Cl)( 5- OCHzCHF?>( S) CH( OAc) C( 0) OH ( a) and Ph( 3- Cn( 5- OCH7CHF9)-(/ g) CH( OH) C(Q)OH (b)

En løsning av Ph(3-Cl)(5-OCH2CHF2)-(/?,S)CH(OH)C(0)OH (3,90 g, 14,6 mmol; se trinn (iv) ovenfor) og Lipase PS "Amano" (2,50 g) i vinylacetat (140 ml) og MTBE (140 ml) ble oppvarmet ved 70°C under nitrogen i 40 timer. Reaksjonsblandingen ble avkjølt til romtemperatur, enzymet fjernet ved filtrering vasking med EtOAc og filtratet konsentrert i vakuum. Kromatografi på silikagel under eluering med CHCl3:MeOH:Et3N (92:6:2) ga trietylaminsaltet av sub-tittelforbindelse (a) som en gul olje. I tillegg, ble trietylaminsaltet av sub-tittelforbindelse (b) (1,47 g) oppnådd og saltet ble oppløst i H20 (100 ml), surgjort med 2 N HC1 og ekstrahert med EtOAc (2 x 75 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilket gir sub-tittelforbindelse (b) (1,00 A solution of Ph(3-Cl)(5-OCH2CHF2)-(/?,S)CH(OH)C(0)OH (3.90 g, 14.6 mmol; see step (iv) above) and Lipase PS "Amano" (2.50 g) in vinyl acetate (140 mL) and MTBE (140 mL) was heated at 70°C under nitrogen for 40 hours. The reaction mixture was cooled to room temperature, the enzyme removed by filtration, washing with EtOAc and the filtrate concentrated in vacuo. Chromatography on silica gel eluting with CHCl3:MeOH:Et3N (92:6:2) gave the triethylamine salt of sub-title compound (a) as a yellow oil. In addition, the triethylamine salt of sub-title compound (b) (1.47 g) was obtained and the salt was dissolved in H 2 O (100 mL), acidified with 2 N HCl and extracted with EtOAc (2 x 75 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give sub-title compound (b) (1.00

g) som et gråhvitt, fast stoff. g) as a grey-white solid.

Data for sub-tittelforbindelse (b): Data for subtitle connection (b):

Sm.p.: 103-106°C Melting point: 103-106°C

Rf = 0,39 (90:8:2 CHCl3:MeOH:Et3N) Rf = 0.39 (90:8:2 CHCl3:MeOH:Et3N)

'H NMR (300 MHz, CD3OD) A 7,13 (s, 1H), 7,04 (s, 1H), 6,97 (s, 1H), 6,17 (tt, J = 4 Hz, J = 55 Hz, 1H), 5,12 (s, 1H), 4,24 (dt, J = 4 Hz, J = 8 Hz, 2H). 1H NMR (300 MHz, CD3OD) Δ 7.13 (s, 1H), 7.04 (s, 1H), 6.97 (s, 1H), 6.17 (tt, J = 4 Hz, J = 55 Hz, 1H), 5.12 (s, 1H), 4.24 (dt, J = 4 Hz, J = 8 Hz, 2H).

<13>C NMR (75 MHz, CD3OD) A 175,5,160,3,144,5, 136,1,121,3,115,7,115,3, (t, J = 240 Hz), 112,9, 73,4, 68,6 (t, J = 29 Hz) <13>C NMR (75 MHz, CD3OD) A 175.5,160.3,144.5, 136.1,121.3,115.7,115.3, (t, J = 240 Hz), 112.9, 73.4, 68.6 (t, J = 29 Hz)

HPLC Analyse: 96,2%, >95,0% ee, ChiralPak AD Kolonne (95:5:0,5 Heks:EtOH:TFA mobil fase) HPLC Analysis: 96.2%, >95.0% ee, ChiralPak AD Column (95:5:0.5 Hex:EtOH:TFA mobile phase)

[I]<25>d = -84,0° (c = 0,85 MeOH) [I]<25>d = -84.0° (c = 0.85 MeOH)

APCI-MS: (M - 1) = 265 m/z APCI-MS: (M - 1) = 265 m/z

(vi) Ph( 3- Cl)( 5- OCH?CHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( Teoc) (vi) Ph( 3- Cl)( 5- OCH?CHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( Teoc)

Til en løsning av Ph(3-Cl)(5-OCH2CHF2)-(Æ)CH(OH)C(0)OH (0,35 g, 1,3 mmol; se trinn (v) ovenfor (forbindelse (b))) i DMF (18 ml) under nitrogen ble tilsatt H-Aze-Pab(Teoc) x HC1 (0,76 g, 1,7 mmol), PyBOP (0,75 g, 1,4 mmol) og DIPEA (0,43 g, 3,3 mmol) ved 0°C. Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur i 20 timer. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCtøEtOH (10:1) og deretter med EtOAc:EtOH (10:1), hvilker gir sub-tittelforbindelsen (0,69 g, 84%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH2CHF2)-(Æ)CH(OH)C(0)OH (0.35 g, 1.3 mmol; see step (v) above (compound (b) )) in DMF (18 mL) under nitrogen was added H-Aze-Pab(Teoc) x HCl (0.76 g, 1.7 mmol), PyBOP (0.75 g, 1.4 mmol) and DIPEA (0 .43 g, 3.3 mmol) at 0°C. The reaction mixture was stirred at 0°C for 2 hours and then at room temperature for 20 hours. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl2EtOH (10:1) and then with EtOAc:EtOH (10:1) to give the sub-title compound (0.69 g, 84%) as a breakable white foam.

Sm.p.: 108-118°C Melting point: 108-118°C

Rf = 0,48 (10:1 CHCl3:EtOH) Rf = 0.48 (10:1 CHCl3:EtOH)

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) A 7,78-7,81 (m, 2H), 7,40-7,43 (m, 2H), 7,09-7,12 (m, 1H), 6,96-7,02 (m, 2H), 6,16 (t, J= 57 Hz, 1H), 5,09-5,20 og 4,75-4,80 (m, 2H), 3,95-4,55 (m, 8H), 2,10-2,75 (m, 2H), 1,04-1,11 (m, 2H), 0,07 (s, 9H) APCI-MS: (M+l) = 625m/z 1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) Δ 7.78-7.81 (m, 2H), 7.40-7.43 (m, 2H), 7.09-7.12 (m , 1H), 6.96-7.02 (m, 2H), 6.16 (t, J= 57 Hz, 1H), 5.09-5.20 and 4.75-4.80 (m, 2H ), 3.95-4.55 (m, 8H), 2.10-2.75 (m, 2H), 1.04-1.11 (m, 2H), 0.07 (s, 9H) APCI - MS: (M+1) = 625m/z

(vii) Ph( 3- Cn( 5- OCHzCHF?)-( j?) CH( OH) C( 0)- Aze- Pab x TFA (vii) Ph( 3- Cn( 5- OCHzCHF?)-( j?) CH( OH) C( 0)- Aze- Pab x TFA

Ph(3-Cl)(5-OCH2CHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,086 g, 0,138 mmol; se trinn (vi) ovenfor), ble oppløst i 3 ml TFA og-fikk reagere i 1 time. TFA ble inndampet og residuet ble fryse tørket fra vann/acetonitril, hvilket gir 0,080 g (98%) av tittelforbindelsen som dens TFA salt. Ph(3-Cl)(5-OCH2CHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.086 g, 0.138 mmol; see step (vi) above), was dissolved in 3 ml TFA and allowed to react for 1 hour. The TFA was evaporated and the residue was freeze dried from water/acetonitrile to give 0.080 g (98%) of the title compound as its TFA salt.

'H-NMR (300 MHz; CD3OD) rotamerer: 8 7,8-7,7 (m, 2H), 7,6-7,5 (m, 2H), 7,15-6,95 (m, 3H), 6,35-5,95 (m, 1H), 5,20 (m, 1H, mindre rotamer), 5,14 (s, 1H, hoved rotamer), 5,10 (s, 1H, mindre rotamer), 4,80 (m, 1H, hoved rotamer), 4,6-4,0 (m, 6H), 2,70 (m, 1H, mindre rotamer), 2,53 (m, 1H, hoved rotamer), 2,29 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer). 1H-NMR (300 MHz; CD 3 OD) rotamers: δ 7.8-7.7 (m, 2H), 7.6-7.5 (m, 2H), 7.15-6.95 (m, 3H ), 6.35-5.95 (m, 1H), 5.20 (m, 1H, minor rotamer), 5.14 (s, 1H, major rotamer), 5.10 (s, 1H, minor rotamer) , 4.80 (m, 1H, major rotamer), 4.6-4.0 (m, 6H), 2.70 (m, 1H, minor rotamer), 2.53 (m, 1H, major rotamer), 2.29 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer).

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 174,0,173,8,173,4,172,9,168,2 <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 174,0,173,8,173,4,172,9,168,2

MS (m/z) 481,2 (M+l)<+>MS (m/z) 481.2 (M+1)<+>

Eksempel 18 Example 18

Ph( 3- Cl)( 5- OCH7CHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCH7CHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe)

Til en løsning av Ph(3-Cl)(5-OCH2CHF2)-(Æ)CH(OH)C(0)OH (0,30 g, 1,7 mmol; se Eksempel 17(v) ovenfor (forbindelse (b))) i DMF (15 ml) under nitrogen ble tilsatt H-AzePab(OMe) x 2HC1 (0,49 g, 1,5 mmol), PyBOP (0,65 g, 1,2 mmol) og DIPEA (0,36 g, 2,8 mmol) ved 0°C. Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur i 20 timer. Blandingen ble konsentrert 1 vakuum og residuet kromatografert tre ganger på silikagel, under eluering først med CHCl3:EtOH (10:1), deretter med EtOAc:EtOH (10:1) og til slutt med CHCl3:MeOH (20:1), hvilket gir tittelforbindelsen (0,47 g, 81%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH2CHF2)-(Æ)CH(OH)C(0)OH (0.30 g, 1.7 mmol; see Example 17(v) above (compound (b) ))) in DMF (15 mL) under nitrogen was added H-AzePab(OMe) x 2HCl (0.49 g, 1.5 mmol), PyBOP (0.65 g, 1.2 mmol) and DIPEA (0, 36 g, 2.8 mmol) at 0°C. The reaction mixture was stirred at 0°C for 2 hours and then at room temperature for 20 hours. The mixture was concentrated in vacuo and the residue chromatographed three times on silica gel, eluting first with CHCl3:EtOH (10:1), then with EtOAc:EtOH (10:1) and finally with CHCl3:MeOH (20:1), which gives the title compound (0.47 g, 81%) as a crushable white foam.

Sm.p.: 65-75°C Melting point: 65-75°C

Rf = 0,37 (10:1 CHCl3:EtOH) Rf = 0.37 (10:1 CHCl3:EtOH)

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) A 7,58-7,60 (m, 2H), 7,32-7,35 (m, 2H), 7,09-7,12 (m, 1H), 6,96-7,02 (m, 2H), 6,16 (t, /= 55 Hz, 1H), 5,08-5,18 og 4,74-4,80 (m, 2H), 3,96-4,50 (m, 6H), 3,80 (s, 3H), 2,10-2,75 (m, 2H) 1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) Δ 7.58-7.60 (m, 2H), 7.32-7.35 (m, 2H), 7.09-7.12 (m , 1H), 6.96-7.02 (m, 2H), 6.16 (t, /= 55 Hz, 1H), 5.08-5.18 and 4.74-4.80 (m, 2H ), 3.96-4.50 (m, 6H), 3.80 (s, 3H), 2.10-2.75 (m, 2H)

APCI-MS: (M+l) = 511m/z. APCI-MS: (M+1) = 511 m/z.

Eksempel 19 Example 19

Ph( 3- Cl)( 5- OCHzF)-( Æ) CH( OH) C( 0)- Aze- Pab x TFA Ph( 3- Cl)( 5- OCHzF)-( Æ) CH( OH) C( 0)- Aze- Pab x TFA

(i) Ph( 3- Cl)( 5- TMSOW/ ?. S) CH( OTMS) CN (i) Ph( 3- Cl)( 5- TMSOW/ ?. S) CH( OTMS) CN

Til en løsning av 3-klor-5-hydroksybenzaldehyd (9,8 g, 62,6 mmol; se Eksempel l(ii) ovenfor) og Znl2 (5,0 g, 15,7 mmol) i vannfri CH2C12 (500 ml) ved 0°C ble tilsatt trimetylsilylcyanid (13,7 g, 138 mmol). Reaksjonsblandingen fikk oppvarmes til romtemperatur og omrørt natten over. Vann (250 ml) ble tilsatt og lagene ble separert. Det vandige laget ble ekstrahert med CH2C12 (2 x 300 ml). De samlede organiske ekstrakter ble tørket (Na2SC«4), filtrert og konsentrert / vakuum, hvilker gir sub-tittelforbindelsen (16,9 g, 83%) som en gul olje som ble anvendt uten ytterligere rensning. To a solution of 3-chloro-5-hydroxybenzaldehyde (9.8 g, 62.6 mmol; see Example 1(ii) above) and Zn12 (5.0 g, 15.7 mmol) in anhydrous CH2 Cl2 (500 mL) at 0°C trimethylsilyl cyanide (13.7 g, 138 mmol) was added. The reaction mixture was allowed to warm to room temperature and stirred overnight. Water (250 mL) was added and the layers were separated. The aqueous layer was extracted with CH 2 Cl 2 (2 x 300 mL). The combined organic extracts were dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give the sub-title compound (16.9 g, 83%) as a yellow oil which was used without further purification.

Rf = 0,42 (3:1 Heks:EtOAc) Rf = 0.42 (3:1 Hex:EtOAc)

'H NMR (300 MHz, CDC13) 8 7,06 (s, 1H), 6,86 (s, 2H), 5,40 (s, 1H), 0,30 (s, 9 H), 0,24 (s,9H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.06 (s, 1H), 6.86 (s, 2H), 5.40 (s, 1H), 0.30 (s, 9H), 0.24 (p. 9H).

(ii) Ph( 3- Cl)( 5- OH)- r/ g. 5JCH( OH) C( Q) OH (ii) Ph( 3- Cl)( 5- OH)- r/ g. 5JCH( OH) C( Q) OH

En løsning av Ph(3-Cl)(5-OTMS)-(R,S)CH(OTMS)CN (22,6 g, 68,8 mmol; se trinn (i) ovenfor) i konsentrert HC1 (200 ml) ble tilbakeløpskokt under nitrogen i 3 timer. Reaksjonsblandingen ble avkjølt til 0°C og gjort basisk langsomt med 2N NaOH. Blandingen ble vasket med Et20 (3 x 100 ml) for å fjerne organisk urenheter. Det vandige laget ble surgjort med 2N HC1 og ekstrahert med EtOAc (3 x 200 ml). De samlede organiske ekstrakter ble tørket (Na2SC«4), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (9,3 g, 67%) som en brun olje som ble anvendt uten ytterligere rensning. A solution of Ph(3-Cl)(5-OTMS)-(R,S)CH(OTMS)CN (22.6 g, 68.8 mmol; see step (i) above) in concentrated HCl (200 mL) was refluxed under nitrogen for 3 hours. The reaction mixture was cooled to 0°C and basified slowly with 2N NaOH. The mixture was washed with Et 2 O (3 x 100 mL) to remove organic impurities. The aqueous layer was acidified with 2N HCl and extracted with EtOAc (3 x 200 mL). The combined organic extracts were dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give the sub-title compound (9.3 g, 67%) as a brown oil which was used without further purification.

Rf = 0,23 (6:3:1 CHCl3:MeOH:konsentrert NH4OH) Rf = 0.23 (6:3:1 CHCl3:MeOH:conc NH4OH)

'H NMR (300 MHz, CD3OD) 8 7,05 (s, 1H), 6,94 (s, 1H), 6,73 (s, 1H), 5,03 (s, 1H). 1 H NMR (300 MHz, CD 3 OD) δ 7.05 (s, 1H), 6.94 (s, 1H), 6.73 (s, 1H), 5.03 (s, 1H).

(iii) Ph( 3- Cl)( 5- OHW/ ?. S) CH( OH) C( 0) OEt (iii) Ph( 3- Cl)( 5- OHW/ ?. S) CH( OH) C( 0) OEt

Til en løsning av Ph(3-Cl)(5-OH)-(Æ,S)CH(OH)C(0)OH (9,3 g, 46,0 mmol; se trinn (ii) ovenfor) i absolutt EtOH (200 ml) ble det tilsatt konsentrert svovelsyre (0,25 ml) og reaksjonen ble tilbakeløpskokt under nitrogen i 4 timer. Reaksjonsblandingen ble avkjølt til 0°C og fast stoff NaHC03 (0,2 g) ble tilsatt. Reaksjonen ble konsentrert 1 vakuum og fordelt med mettet NaHC03 (100 ml) og Et20 (3 x 50 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (6,9 g, 65%) som en gul olje som ble anvendt uten ytterligere rensning. To a solution of Ph(3-Cl)(5-OH)-(Æ,S)CH(OH)C(0)OH (9.3 g, 46.0 mmol; see step (ii) above) in absolute To EtOH (200 mL) was added concentrated sulfuric acid (0.25 mL) and the reaction was refluxed under nitrogen for 4 h. The reaction mixture was cooled to 0°C and solid NaHCO 3 (0.2 g) was added. The reaction was concentrated in vacuo and partitioned with saturated NaHCO 3 (100 mL) and Et 2 O (3 x 50 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (6.9 g, 65%) as a yellow oil which was used without further purification.

Rf = 0,62 (6:3:1 CHCl3:MeOH:konsentrert NH4OH). Rf = 0.62 (6:3:1 CHCl 3 :MeOH:conc NH 4 OH).

'H NMR (300 MHz, CDC13) 8 6,99 (s, 1H), 6,81 (s, 2H), 5,07 (s, 1H), 4,16-4,32 (m, 2H), 1,23 (t,7=7Hz, 3H). 1 H NMR (300 MHz, CDCl 3 ) δ 6.99 (s, 1H), 6.81 (s, 2H), 5.07 (s, 1H), 4.16-4.32 (m, 2H), 1.23 (t.7=7Hz, 3H).

(iv) Ph( 3- Cl)( 5- OCH9F)- r/ ?. 5) CH( OH) C( Q) OEt (iv) Ph( 3- Cl)( 5- OCH9F)- r/ ?. 5) CH(OH)C(Q)OEt

Til en løsning av Ph(3-Cl)(5-OH)-(/?,S)CH(OH)C(0)OEt (6,1 g, 26,8 mmol; se trinn (iii) ovenfor) i DMF (100 ml) i en forseglet kolbe under nitrogen ved 0°C ble det tilsatt cesiumkarbonat (13,1 g, 40,2 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 15 minutter fulgt av tilsetning av kaliumjodid (0,5 g, 2,7 mmol). Reaksjonsblandingen ble avkjølt til -78°C og klorfluormetan (18,4 g, 268 mmol) ble boblet inn i karet. Den forseglete kolben fikk deretter oppvarmes til romtemperatur og omrørt i 18 timer. Reaksjonsblandingen ble avkjølt til 0°C, luftet forsiktig for å fjerne eventuelt overskudd av klorfluormetan og fordelt med H2O (20 ml) og EtiO (3 x 50 ml). De samlede organiske lag ble vasket med saltvann (2 x 50 ml), tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med Heks:EtOAc (gradient fra 9:1 til 3:1) ga sub-tittelforbindelsen (2,4 g, 35%) som en lysegul olje. Anmerkning: Forbindelsen er så vidt uv-synlig på TLC. Den kan visualiseres ved å merke TLC med bromcresol grønn. To a solution of Ph(3-Cl)(5-OH)-(/?,S)CH(OH)C(0)OEt (6.1 g, 26.8 mmol; see step (iii) above) in To DMF (100 mL) in a sealed flask under nitrogen at 0°C was added cesium carbonate (13.1 g, 40.2 mmol). The reaction mixture was stirred at 0°C for 15 min followed by the addition of potassium iodide (0.5 g, 2.7 mmol). The reaction mixture was cooled to -78°C and chlorofluoromethane (18.4 g, 268 mmol) was bubbled into the vessel. The sealed flask was then allowed to warm to room temperature and stirred for 18 hours. The reaction mixture was cooled to 0°C, gently aerated to remove any excess chlorofluoromethane and partitioned with H 2 O (20 mL) and EtiO (3 x 50 mL). The combined organic layers were washed with brine (2 x 50 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (gradient from 9:1 to 3:1) afforded the sub-title compound (2.4 g, 35%) as a pale yellow oil. Note: The compound is barely visible on TLC. It can be visualized by labeling TLC with bromocresol green.

Rf = 0,46 (2:1 Heks:EtOAc) Rf = 0.46 (2:1 Hex:EtOAc)

<*>H NMR (300 MHz, CDC13) 8 7,21 (s, 1H), 7,08 (s, 1H), 7,05 (s, 1H), 5,70 (d, 7H.F = 54 Hz, 2H), 5,12 (d, 7 = 5 Hz, 1H), 3,80-4,35 (m, 2H), 3,50 (d, 7 = 5 Hz, 1H), 1,26 (t, 7 = 7 Hz, 3H). <*>H NMR (300 MHz, CDCl 3 ) δ 7.21 (s, 1H), 7.08 (s, 1H), 7.05 (s, 1H), 5.70 (d, 7H.F = 54 Hz, 2H), 5.12 (d, 7 = 5 Hz, 1H), 3.80-4.35 (m, 2H), 3.50 (d, 7 = 5 Hz, 1H), 1.26 ( t, 7 = 7 Hz, 3H).

(v) Ph( 3- Cl)( 5- OCH2FW/ g. S) CH( OH) C( 0) OH (v) Ph( 3- Cl)( 5- OCH2FW/ g. S) CH( OH) C( 0) OH

Til en løsning av Ph(3-Cl)(5-OCH2F)-(/?,5)CH(OH)C(0)OEt (1,8 g, 6,8 mmol; se trinn (iv) ovenfor) i H20:THF (30 ml, 1:2) ved 0°C under nitrogen ble det tilsatt litiumhydroksydmonohydrat (0,40 g, 10,3 mmol). Blandingen ble omrørt ved 0°C i 2 timer. Reaksjonsblandingen ble konsentrert i vakuum og fordelt med H2O (5 ml) og Et20 (2 x 20 ml). Det vandige laget ble surgjort forsiktig med 0,2N HC1 ved 0°C og ekstrahert med EtOAc (3 x 30 ml). De samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (1,4 g, 87%) som en fargeløs olje som stivnet til et hvitt, fast stoff ved henstand. To a solution of Ph(3-Cl)(5-OCH2F)-(/?,5)CH(OH)C(0)OEt (1.8 g, 6.8 mmol; see step (iv) above) in H 2 O:THF (30 mL, 1:2) at 0°C under nitrogen was added lithium hydroxide monohydrate (0.40 g, 10.3 mmol). The mixture was stirred at 0°C for 2 hours. The reaction mixture was concentrated in vacuo and partitioned with H 2 O (5 mL) and Et 2 O (2 x 20 mL). The aqueous layer was acidified gently with 0.2N HCl at 0°C and extracted with EtOAc (3 x 30 mL). The combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (1.4 g, 87%) as a colorless oil which solidified to a white solid on standing.

Rf = 0,43 (6:2:1 CHCl3:MeOH:Et3N) Rf = 0.43 (6:2:1 CHCl3:MeOH:Et3N)

<l>H NMR (300 MHz, CD3OD) 8 7,24 (s, 1H), 7,17 (s, 1H), 7,07 (s, 1H), 5,78 (d, 7H-f = 54 Hz, 2H), 5,13 (s, 1H). <1>H NMR (300 MHz, CD3OD) δ 7.24 (s, 1H), 7.17 (s, 1H), 7.07 (s, 1H), 5.78 (d, 7H-f = 54 Hz, 2H), 5.13 (s, 1H).

(vi) Ph( 3- Cl)( 5- OCH2F)-(/ ?) CH( OH) C( 0) OH ( a) oe Ph( 3- Cl)( 5- OCH2F)-( S) CH( OAc) C( Q) OH ( b) (vi) Ph( 3- Cl)( 5- OCH2F)-(/ ?) CH( OH) C( 0) OH ( a) oe Ph( 3- Cl)( 5- OCH2F)-( S) CH( OAc ) C(Q)OH (b)

En blanding av Ph(3-Cl)(5-OCH2F)-(/?,S)CH(OH)C(0)OH (3,2 g, 13,9 mmol; se trinn (v) ovenfor) og Lipase PS "Amano" (1,9 g) i vinylacetat (150 ml) og MTBE (150 ml) ble oppvarmet ved 70°C under en nitrogen-atmosfære i 3 d. Reaksjonsblandingen ble avkjølt, filtrert gjennom Celite® og filterkaken vasket med EtOAc. Filtratet ble konsentrert i vakuum og underkastet flash kromatografi på en silikagel under eluering med CHCl3:MeOH:Et3N (15:1:0,5), hvilket gir trietylaminsaltet av sub-tittelforbindelse (a) A mixture of Ph(3-Cl)(5-OCH2F)-(/?,S)CH(OH)C(0)OH (3.2 g, 13.9 mmol; see step (v) above) and Lipase PS "Amano" (1.9 g) in vinyl acetate (150 mL) and MTBE (150 mL) was heated at 70°C under a nitrogen atmosphere for 3 d. The reaction mixture was cooled, filtered through Celite® and the filter cake washed with EtOAc . The filtrate was concentrated in vacuo and subjected to flash chromatography on a silica gel eluting with CHCl3:MeOH:Et3N (15:1:0.5) to give the triethylamine salt of sub-title compound (a)

(0,50 g, 21%) som ble anvendt uten nøytralisering. I tillegg, ble trietylaminsaltet av sub-tittelforbindelse (b) (0,46 g, 20%) oppnådd. (0.50 g, 21%) which was used without neutralization. In addition, the triethylamine salt of sub-title compound (b) (0.46 g, 20%) was obtained.

Data for Sub- Tittelforbindelse ( a) : Data for Sub-Title connection (a) :

Rf = 0,19 (15:1:0,5 CHCl3:MeOH:Et3N) Rf = 0.19 (15:1:0.5 CHCl3:MeOH:Et3N)

'H NMR (300 MHz, CD3OD) 8 7,26 (s, 1H), 7,18 (s, 1H), 6,97 (s, 1H), 5,74 (d, 7H-f = 54 Hz, 2H), 4,81 (s, 1H), 3,17 (q, / = 7 Hz, 6H), 1,28 (t, / = 7 Hz, 9H). 1 H NMR (300 MHz, CD 3 OD) δ 7.26 (s, 1H), 7.18 (s, 1H), 6.97 (s, 1H), 5.74 (d, 7H-f = 54 Hz, 2H), 4.81 (s, 1H), 3.17 (q, / = 7 Hz, 6H), 1.28 (t, / = 7 Hz, 9H).

Data for Sub- Tittelforbindelse ( b) Data for Sub-Title connection (b)

Rf = 0,33 (15:1:0,5 CHCl3:MeOH:Et3N) Rf = 0.33 (15:1:0.5 CHCl3:MeOH:Et3N)

<*>H NMR (300 MHz, CD3OD) 8 7,28 (s, 1H), 7,19 (s, 1H), 7,09 (s, 1H), 5,76 (d, 7H-f = 54 Hz, 2H), 5,75 (s, 1H), 3,17 (q, J = 7 Hz, 6H), 2,16 (s, 3H), 1,28 (t, J = 7 Hz, 9H). <*>H NMR (300 MHz, CD3OD) δ 7.28 (s, 1H), 7.19 (s, 1H), 7.09 (s, 1H), 5.76 (d, 7H-f = 54 Hz, 2H), 5.75 (s, 1H), 3.17 (q, J = 7 Hz, 6H), 2.16 (s, 3H), 1.28 (t, J = 7 Hz, 9H) .

(vii) Ph( 3- Cl)( 5- OCH7F)-(/ g) CH( OH) C( 0)- Aze- Pab( Teoc) (vii) Ph( 3- Cl)( 5- OCH7F)-(/ g) CH( OH) C( 0)- Aze- Pab( Teoc)

Til en løsning av trietylaminsaltet av Ph(3-Cl)(5-OCH2F)-(/?)CH(OH)C(0)OH (0,50 g, 1,50 mmol; se trinn (vi) ovenfor) og HAze-Pab(Teoc)»HCl (0,87 g, 1,90 mmol) i tørr DMF (15 ml) under nitrogen ved 0°C ble tilsatt PyBOP (0,85 g, 2,60 mmol) og DIPEA (0,48 g, 3,70 mmol). Reaksjonen fikk oppvarmes til romtemperatur og omrørt natten over. Reaksjonsblandingen ble konsentrert i vakuum og flash kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (9:1) og deretter med EtOAc:EtOH (20:1), hvilker gir sub-tittelforbindelsen (0,23 g, 26%) som et knusbart hvitt skum. To a solution of the triethylamine salt of Ph(3-Cl)(5-OCH2F)-(/?)CH(OH)C(0)OH (0.50 g, 1.50 mmol; see step (vi) above) and HAze-Pab(Teoc)»HCl (0.87 g, 1.90 mmol) in dry DMF (15 mL) under nitrogen at 0 °C was added PyBOP (0.85 g, 2.60 mmol) and DIPEA (0 .48 g, 3.70 mmol). The reaction was allowed to warm to room temperature and stirred overnight. The reaction mixture was concentrated in vacuo and flash chromatographed twice on silica gel, eluting first with CHCl 3 :EtOH (9:1) and then with EtOAc:EtOH (20:1) to give the sub-title compound (0.23 g, 26% ) as a crushable white foam.

Sm.p.: 88-92°C Melting point: 88-92°C

Rf = 0,61 (9:1 CHCl3:EtOH) Rf = 0.61 (9:1 CHCl3:EtOH)

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,81 (d, J = 8 Hz, 2H), 7,40-7,42 (m, 2H), 7,06-7,23 (m, 3H), 5,76 (d, 7H-f = 51 Hz, 2H), 5,10-5,16 og 4,77-4,83 (m, 2H), 3,80-4,49 (m, 6H), 2,30-2,53 (m, 2H), 1,08 (t, / = 7 Hz, 2H), 0,08(s, 9H). APCI-MS (M + 1) = 593 m/z 1H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.81 (d, J = 8 Hz, 2H), 7.40-7.42 (m, 2H), 7.06-7.23 ( m, 3H), 5.76 (d, 7H-f = 51 Hz, 2H), 5.10-5.16 and 4.77-4.83 (m, 2H), 3.80-4.49 ( m, 6H), 2.30-2.53 (m, 2H), 1.08 (t, / = 7 Hz, 2H), 0.08(s, 9H). APCI-MS (M + 1) = 593 m/z

(viii) Ph( 3- Cl¥5- OCH?F)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA (viii) Ph( 3- Cl¥5- OCH?F)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA

Ph(3-Cl)(5-OCH2F)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,051 g, 0,086 mmol; se trinn (vii) ovenfor), ble oppløst i 3 ml TFA og fikk reagere i 20 min. TFA ble inndampet og residuet ble fryse tørket fra vann/acetonitril. Produktet var 95% rent med 5% av defluormetylert materiale. Forsøk på å rense det ved preparativ RPLC med CH3CN:0,1M NH4OAC sviktet og materialet, delvis som et acetat, ble oppløst i 5 ml TFA, inndampet og fryse tørket, hvilket gir 26 mg (51%) av tittelforbindelsen som dens TFA salt. Renhet: 95%. Ph(3-Cl)(5-OCH2F)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.051 g, 0.086 mmol; see step (vii) above), was dissolved in 3 ml TFA and allowed to react for 20 min. The TFA was evaporated and the residue was freeze dried from water/acetonitrile. The product was 95% pure with 5% defluoromethylated material. Attempts to purify it by preparative RPLC with CH3CN:0.1M NH4OAC failed and the material, partly as an acetate, was dissolved in 5 mL of TFA, evaporated and freeze dried, yielding 26 mg (51%) of the title compound as its TFA salt . Purity: 95%.

'H-NMR (600 MHz; CD3OD) rotamerer: 8 7,8-7,7 (m, 2H), 7,6-7,5 (m, 2H), 7,21 (s, 1H, hoved rotamer), 7,17 (s, 1H, mindre rotamer), 7,13 (s, 1H, hoved rotamer), 7,09 (s, 1H, mindre rotamer), 7,07 (m, 1H, hoved rotamer), 7,04 (m, 1H, mindre rotamer), 5,73 (d, 2H), 5.18 (m, 1H, mindre rotamer), 5,16 (s, 1H, hoved rotamer), 5,09 (s, 1H, mindre rotamer), 4,78 (m, 1H, mindre rotamer), 4,56 (d, 1H, hoved rotamer), 4,50 (d, 1H, mindre rotamer), 4,46 (d, 1H, mindre rotamer), 4,45 (d, 1H, hoved rotamer), 4,35 (m, 1H, hoved rotamer), 4,14 (m, 1H, hoved rotamer), 4,05 (m, 1H, mindre rotamer), 3,97 (m, 1H, mindre rotamer), 2,68 (m, 1H, mindre rotamer), 2,52 (m, 1H, hoved rotamer), 2,28 (m, 1H, hoved rotamer), 2.19 (m, 1H, mindre rotamer). 1H-NMR (600 MHz; CD3OD) rotamers: δ 7.8-7.7 (m, 2H), 7.6-7.5 (m, 2H), 7.21 (s, 1H, main rotamer) , 7.17 (s, 1H, minor rotamer), 7.13 (s, 1H, major rotamer), 7.09 (s, 1H, minor rotamer), 7.07 (m, 1H, major rotamer), 7 .04 (m, 1H, minor rotamer), 5.73 (d, 2H), 5.18 (m, 1H, minor rotamer), 5.16 (s, 1H, major rotamer), 5.09 (s, 1H, minor rotamer), 4.78 (m, 1H, minor rotamer), 4.56 (d, 1H, major rotamer), 4.50 (d, 1H, minor rotamer), 4.46 (d, 1H, minor rotamer ), 4.45 (d, 1H, major rotamer), 4.35 (m, 1H, major rotamer), 4.14 (m, 1H, major rotamer), 4.05 (m, 1H, minor rotamer), 3.97 (m, 1H, minor rotamer), 2.68 (m, 1H, minor rotamer), 2.52 (m, 1H, major rotamer), 2.28 (m, 1H, major rotamer), 2.19 ( m, 1H, minor rotamer).

<13>C-NMR (150 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 173,9,173,3,172,9,168,2. <13>C-NMR (150 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 173,9,173,3,172,9,168,2.

ESI-MS+: (M+l) = 449 (m/z) ESI-MS+: (M+1) = 449 (m/z)

Eksempel 20 Example 20

Ph( 3- Cl)( 5- OCH9F)-(/ g) CH( OH) C( Q)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCH9F)-(/ g) CH( OH) C( Q)- Aze- Pab( OMe)

Til en løsning av trietylaminsaltet av Ph(3-Cl)(5-OCH2F)-(Æ)CH(OH)C(0)OH (0,60 g, 1,80 mmol; se Eksempel 19(vi)) og HAze-Pab(OMe)»2HCl (0,79 g, 2,30 mmol) i DMF (15 ml) under nitrogen ved 0°C ble tilsatt PyBOP (1,04 g, 1,90 mmol) og DIPEA (0,58 g, 4,50 mmol). Reaksjonen fikk oppvarmes til romtemperatur og omrørt natten over. Reaksjonsblandingen ble konsentrert i vakuum og flash kromatografert tre ganger på silikagel, under eluering først med CHCtøEtOH (9:1) og deretter to ganger med EtOAc:EtOH (20:1), hvilket gir tittelforbindelsen (0,22 g, 26%) som et knusbart hvitt skum. To a solution of the triethylamine salt of Ph(3-Cl)(5-OCH2F)-(Æ)CH(OH)C(0)OH (0.60 g, 1.80 mmol; see Example 19(vi)) and HAze -Pab(OMe)»2HCl (0.79 g, 2.30 mmol) in DMF (15 mL) under nitrogen at 0 °C was added PyBOP (1.04 g, 1.90 mmol) and DIPEA (0.58 g, 4.50 mmol). The reaction was allowed to warm to room temperature and stirred overnight. The reaction mixture was concentrated in vacuo and flash chromatographed three times on silica gel, eluting first with CHCl2EtOH (9:1) and then twice with EtOAc:EtOH (20:1) to give the title compound (0.22 g, 26%) as a breakable white foam.

Sm.p.: 66-70°C Melting point: 66-70°C

Rf = 0,45 (9:1 CHCl3:EtOH) Rf = 0.45 (9:1 CHCl3:EtOH)

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,59 (d, / = 8 Hz, 2H), 7,32 (d, J = 7 Hz, 2H), 7,06-7,23 (m, 3H), 5,75 (s, 7H-f = 54 Hz, 1H), 5,10-5,16 og 4,78-4,84 (m, 2H), 4,11-4,45 (m, 4H), 3,80 (s, 3H), 2,10-2,75 (m, 2H). 1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) δ 7.59 (d, / = 8 Hz, 2H), 7.32 (d, J = 7 Hz, 2H), 7.06-7.23 (m, 3H), 5.75 (s, 7H-f = 54 Hz, 1H), 5.10-5.16 and 4.78-4.84 (m, 2H), 4.11-4.45 (m, 4H), 3.80 (s, 3H), 2.10-2.75 (m, 2H).

<13>C-NMR (150 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 173,0,170,8,170,7,152,5. <13>C-NMR (150 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 173.0.170.8.170.7.152.5.

APCI-MS:(M + 1) = 479 m/z APCI-MS: (M + 1) = 479 m/z

Eksempel 21 Example 21

Ph( 3- Cn( 5- OCH2CH2F)-(/ ?) CH( OH) C( 0)- Aze- Pab x TFA Ph( 3- Cn( 5- OCH2CH2F)-(/ ?) CH( OH) C( 0)- Aze- Pab x TFA

(i) ( 2- Monolfuoretvl) metansulfonat (i) (2-Monolfluoroethyl)methanesulfonate

Til en magnetisk omrørt løsning av 2-fluoretanol (5,0 g, 78,0 mmol) i CH2CI2 (90 ml) under nitrogen ved 0°C ble tilsatt trietylamin (23,7 g, 234 mmol) og metansulfonylklorid (10,7 g, 93,7 mmol). Blandingen ble omrørt ved 0°C i 1,5 timer, fortynnet med CH2CI2 (100 ml) og vasket med 2N HC1 (100 ml). Det vandige laget ble ekstrahert med CH2C12 (50 ml) og de samlede organiske ekstrakter vasket med saltvann (75 ml), tørket (Na2SC«4), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (9,7 g, 88%) som en gul olje som ble anvendt uten ytterligere rensning. To a magnetically stirred solution of 2-fluoroethanol (5.0 g, 78.0 mmol) in CH 2 Cl 2 (90 mL) under nitrogen at 0 °C was added triethylamine (23.7 g, 234 mmol) and methanesulfonyl chloride (10.7 g, 93.7 mmol). The mixture was stirred at 0°C for 1.5 h, diluted with CH 2 Cl 2 (100 mL) and washed with 2N HCl (100 mL). The aqueous layer was extracted with CH 2 Cl 2 (50 mL) and the combined organic extracts washed with brine (75 mL), dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give the sub-title compound (9.7 g, 88% ) as a yellow oil which was used without further purification.

'H NMR (300 MHz, CDCI3) 8 4,76 (t, J = 4 Hz, 1H), 4,64 (t, J = 4 Hz, 1H), 4,52 (t, J = 4 Hz, 1H), 4,43 (t, / = 4 Hz, 1H), 3,09 (s, 3H). 1H NMR (300 MHz, CDCl 3 ) δ 4.76 (t, J = 4 Hz, 1H), 4.64 (t, J = 4 Hz, 1H), 4.52 (t, J = 4 Hz, 1H ), 4.43 (t, / = 4 Hz, 1H), 3.09 (s, 3H).

(ii) 3- klor- 5- monofluoretoksvbenzaldehvd (ii) 3- chloro- 5- monofluoroethoxybenzaldehvd

Til en løsning av 3-klor-5-hydroksybenzaldehyd (8,2 g, 52,5 mmol; se Eksempel l(ii) ovenfor) og kaliumkarbonat (9,4 g, 68,2 mmol) i DMF (10 ml) under nitrogen ble det tilsatt en løsning av (2-monofluoretyl) metansulfonat (9,7 g, 68,2 mmol; se trinn (i) ovenfor) i DMF (120 ml) dråpevis ved romtemperatur. Blandingen ble oppvarmet til 100°C i 5 timer og deretter omrørt natten over ved romtemperatur. Reaksjonsblandingen ble avkjølt til 0°C, hellet i is-kald 2N HC1 og ekstrahert med EtOAc. De samlede organiske ekstrakter ble vasket med saltvann, tørket (Na2S04), filtrert og konsentrert i vakuum. Den brune oljen ble kromatografert på silikagel under eluering med Heks:EtOAc (4:1), hvilker gir sub-tittelforbindelsen (7,6 g, 71%) som en gul olje. To a solution of 3-chloro-5-hydroxybenzaldehyde (8.2 g, 52.5 mmol; see Example 1(ii) above) and potassium carbonate (9.4 g, 68.2 mmol) in DMF (10 mL) under nitrogen, a solution of (2-monofluoroethyl) methanesulfonate (9.7 g, 68.2 mmol; see step (i) above) in DMF (120 mL) was added dropwise at room temperature. The mixture was heated to 100°C for 5 hours and then stirred overnight at room temperature. The reaction mixture was cooled to 0°C, poured into ice-cold 2N HCl and extracted with EtOAc. The combined organic extracts were washed with brine, dried (Na 2 SO 4 ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with Hex:EtOAc (4:1) to give the sub-title compound (7.6 g, 71%) as a yellow oil.

'H NMR (300 MHz, CDCI3) 8 9,92 (s, 1H), 7,48 (s, 1H), 7,32 (s, 1H), 7,21 (s, 1H), 4,87 (t, J = 4 Hz, 1H), 4,71 (t, J = 3 Hz, 1H), 4,33 (t, / = 3 Hz, 1H), 4,24 (t, J = 3 Hz, 1H). 1 H NMR (300 MHz, CDCl 3 ) δ 9.92 (s, 1H), 7.48 (s, 1H), 7.32 (s, 1H), 7.21 (s, 1H), 4.87 ( t, J = 4 Hz, 1H), 4.71 (t, J = 3 Hz, 1H), 4.33 (t, / = 3 Hz, 1H), 4.24 (t, J = 3 Hz, 1H ).

(iii) Ph( 3- Cl)( 5- OCH9CH?F)-(/ g. 5) CH( OTMS) CN (iii) Ph( 3- Cl)( 5- OCH9CH?F)-(/ g. 5) CH( OTMS) CN

Til en løsning av 3-klor-5-monofluoretoksybenzaldehyd (7,6 g, 37,5 mmol; se trinn (ii) ovenfor) og sinkjodid (3,0 g, 9,38 mmol) i CH2CI2 (310 ml) ble tilsatt trimetylsilylcyanid (7,4 g, 75,0 mmol) dråpevis ved 0°C under nitrogen. Blandingen ble omrørt ved 0°C i 3 timer og ved romtemperatur natten over. Reaksjonen ble fortynnet med H2O (300 ml), det organiske laget ble separert, tørket (Na2S04), filtrert og konsentrert / vakuum, hvilker gir sub-tittelforbindelsen (10,6 g, 94%) som en brun olje som ble anvendt uten ytterligere rensning eller karakterisering. To a solution of 3-chloro-5-monofluoroethoxybenzaldehyde (7.6 g, 37.5 mmol; see step (ii) above) and zinc iodide (3.0 g, 9.38 mmol) in CH 2 Cl 2 (310 mL) was added trimethylsilyl cyanide (7.4 g, 75.0 mmol) dropwise at 0 °C under nitrogen. The mixture was stirred at 0°C for 3 hours and at room temperature overnight. The reaction was diluted with H 2 O (300 mL), the organic layer was separated, dried (Na 2 SO 4 ), filtered and concentrated / vacuum to give the sub-title compound (10.6 g, 94%) as a brown oil which was used without further purification or characterization.

(iv) Ph( 3- Cl)( 5- OCH2CH?FW/ g., Sr) CH( OH) C( 0) OH (iv) Ph( 3- Cl)( 5- OCH2CH?FW/ g., Sr) CH( OH) C( 0) OH

Konsentrert saltsyre (100 ml) ble satt til Ph(3-Cl)(5-OCH2CH2F)-(/?,S)CH(OTMS)CN (10,6 g, 5,8 mmol; se trinn (iii) ovenfor) og løsningen omrørt ved 100°C i 3 timer. Etter avkjøling til romtemperatur, ble reaksjonen ytterligere avkjølt til 0°C, gjort basisk langsomt med 3N NaOH (-300 ml) og vasket med Et20 (3 x 200 ml). Det vandige laget ble surgjort med 2N HC1 (80 ml) og ekstrahert med EtOAc (3 x Concentrated hydrochloric acid (100 mL) was added to Ph(3-Cl)(5-OCH2CH2F)-(/?,S)CH(OTMS)CN (10.6 g, 5.8 mmol; see step (iii) above) and the solution stirred at 100°C for 3 hours. After cooling to room temperature, the reaction was further cooled to 0°C, basified slowly with 3N NaOH (-300 mL) and washed with Et 2 O (3 x 200 mL). The aqueous layer was acidified with 2N HCl (80 mL) and extracted with EtOAc (3 x

300 ml). De samlede EtOAc ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (8,6 g, 98%) som et blekgult, fast stoff som ble anvendt uten ytterligere rensning. 300 ml). The combined EtOAc extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (8.6 g, 98%) as a pale yellow solid which was used without further purification.

Rf = 0,28 (90:8:2 CHCl3:MeOH:konsentrert NH4OH) Rf = 0.28 (90:8:2 CHCl3:MeOH:conc NH4OH)

'H NMR (300 MHz, CD3OD) 5 7,09 (s, 1H), 7,02 (s, 1H), 6,93 (s, 1H), 5,11 (s, 1H), 4,77-4,81 (m, 1H), 4,62-4,65 (m, 1H), 4,25-4,28 (m, 1H), 4,15-4,18 (m, 1H). 1 H NMR (300 MHz, CD 3 OD) δ 7.09 (s, 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), 4.77- 4.81 (m, 1H), 4.62-4.65 (m, 1H), 4.25-4.28 (m, 1H), 4.15-4.18 (m, 1H).

(v) Ph( 3- Cl)( 5- OCH?CH?F)-( S) CH( OAc) C( 0) OH ( a) og Ph( 3- Cl)( 5- OCH?CH7F)-(/ ?) CH( OH) C( Q) OH ( b) (v) Ph( 3- Cl)( 5- OCH?CH?F)-( S) CH( OAc) C( 0) OH ( a) and Ph( 3- Cl)( 5- OCH?CH7F)-( / ?) CH( OH) C( Q) OH ( b)

En løsning av Ph(3-Cl)(5-OCH2CH2F)-(/?,5)CH(OH)C(0)OH (8,6 g, 34,5 mmol; se trinn (iv) ovenfor) og Lipase PS "Amano" (4,0 g) i vinylacetat (250 ml) og MTBE (250 ml) ble oppvarmet ved 70°C under nitrogen i 3 d. Reaksjonsblandingen ble avkjølt til romtemperatur og enzymet fjernet ved filtrering gjennom Celite®. Filterkaken ble vasket med EtOAc og filtratet konsentrert i vakuum. Kromatografi på silikagel under eluering med CHCl3:MeOH:Et3N (90:8:2) ga trietylaminsaltet av sub-tittelforbindelse (a) som en gul olje. I tillegg, ble trietylaminsaltet av sub-tittelforbindelse (b) (4,0 g) oppnådd. Saltet av sub-tittelforbindelse (b) ble oppløst i H20 (250 ml), surgjort med 2N HC1 og ekstrahert med EtOAc (3 x 200 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (b) (2,8 g, 32%) som en gul olje. A solution of Ph(3-Cl)(5-OCH2CH2F)-(/?,5)CH(OH)C(0)OH (8.6 g, 34.5 mmol; see step (iv) above) and Lipase PS "Amano" (4.0 g) in vinyl acetate (250 mL) and MTBE (250 mL) was heated at 70°C under nitrogen for 3 d. The reaction mixture was cooled to room temperature and the enzyme removed by filtration through Celite®. The filter cake was washed with EtOAc and the filtrate concentrated in vacuo. Chromatography on silica gel eluting with CHCl3:MeOH:Et3N (90:8:2) gave the triethylamine salt of sub-title compound (a) as a yellow oil. In addition, the triethylamine salt of sub-title compound (b) (4.0 g) was obtained. The salt of sub-title compound (b) was dissolved in H 2 O (250 mL), acidified with 2N HCl and extracted with EtOAc (3 x 200 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (b) (2.8 g, 32%) as a yellow oil.

Data for Sub- Tittelforbindelse ( b) : Data for Sub-Title connection (b) :

Rf = 0,28 (90:8:2 CHCl3:MeOH:konsentrert NH4OH) Rf = 0.28 (90:8:2 CHCl3:MeOH:conc NH4OH)

'H NMR (300 MHz, CD3OD) 8 7,09 (s, 1H), 7,02 (s, 1H), 6,93 (s, 1H), 5,11 (s, 1H), 4,77-4,81 (m, 1H), 4,62-4,65 (m, 1H), 4,25-4,28 (m, 1H), 4,15-4,18 (m, 1H). 1 H NMR (300 MHz, CD 3 OD) δ 7.09 (s, 1H), 7.02 (s, 1H), 6.93 (s, 1H), 5.11 (s, 1H), 4.77- 4.81 (m, 1H), 4.62-4.65 (m, 1H), 4.25-4.28 (m, 1H), 4.15-4.18 (m, 1H).

(vi) Ph( 3- Cl)( 5- OCH7CH7F)-(/ ?) CH( OH) C( 0)- Aze- Pab( Teoc) (vi) Ph( 3- Cl)( 5- OCH7CH7F)-(/ ?) CH( OH) C( 0)- Aze- Pab( Teoc)

Til en løsning av Ph(3-Cl)(5-OCH2CH2F)-(/?)CH(OH)C(0)OH (940 mg, 3,78 mmol; se trinn (v) ovenfor) i DMF (30 ml) under nitrogen ved 0°C ble tilsatt HAze-Pab(Teoc)»HCl (2,21 g, 4,91 mmol), PyBOP (2,16 g, 4,15 mmol) og DIPEA (1,22 g, 9,45 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur i 4 timer. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (15:1) og deretter med EtOAc:EtOH (20:1), hvilker gir sub-tittelforbindelsen (450 mg, 20%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH2CH2F)-(/?)CH(OH)C(0)OH (940 mg, 3.78 mmol; see step (v) above) in DMF (30 mL ) under nitrogen at 0°C was added HAze-Pab(Teoc)»HCl (2.21 g, 4.91 mmol), PyBOP (2.16 g, 4.15 mmol) and DIPEA (1.22 g, 9 .45 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature for 4 hours. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl3:EtOH (15:1) and then with EtOAc:EtOH (20:1) to give the sub-title compound (450 mg, 20%) as a breakable white foam.

Sm.p.: 80-88°C Melting point: 80-88°C

Rf = 0,60 (10:1 CHCl3:EtOH) Rf = 0.60 (10:1 CHCl3:EtOH)

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,79 (d, J = 8 Hz, 2H), 7,42 (d, J = 8 Hz, 2H), 7,05-7,08 (m, 1H), 6,93-6,99 (m, 2H), 5,08-5,13 (m, 1H), 4,75-4,80 (m, 2H), 4,60-4,68 (m, 1H), 3,95-4,55 (m, 8H), 2,10-2,75 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s,9H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.79 (d, J = 8 Hz, 2H), 7.42 (d, J = 8 Hz, 2H), 7.05-7 .08 (m, 1H), 6.93-6.99 (m, 2H), 5.08-5.13 (m, 1H), 4.75-4.80 (m, 2H), 4.60 -4.68 (m, 1H), 3.95-4.55 (m, 8H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0 ,08 (p,9H).

APCI-MS: (M + 1) = 607 m/z. APCI-MS: (M + 1) = 607 m/z.

(vii) Ph( 3- Cl)( 5- OCH7CH?F)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA (vii) Ph( 3- Cl)( 5- OCH7CH?F)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA

Ph(3-Cl)(5-OCH2CH2F)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,357 g, 0,589 mmol; se trinn (vi) ovenfor), ble oppløst i 10 ml TFA og fikk reagere i 40 min. TFA ble inndampet og residuet ble fryse tørket fra vann/acetonitril, hvilket gir 0,33 g (93%) av tittelforbindelsen som dens TFA salt. Ph(3-Cl)(5-OCH2CH2F)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.357 g, 0.589 mmol; see step (vi) above), was dissolved in 10 ml TFA and allowed to react for 40 min. The TFA was evaporated and the residue was freeze dried from water/acetonitrile to give 0.33 g (93%) of the title compound as its TFA salt.

'H-NMR (600 MHz; CD3OD) rotamerer: 8 7,8-7,7 (m, 2H), 7,54 (d, 2H), 7,08 (s, 1H, hoved rotamer), 7,04 (s, 1H, mindre rotamer), 6,99 (s, 1H, hoved rotamer), 6,95 (s, 1H), 6,92 (s, 1H, mindre rotamer), 5,18 (m, 1H, mindre rotamer), 5,14 (s, 1H, hoved rotamer), 5,08 (s, 1H, mindre rotamer), 4,80 (m, 1H, hoved rotamer), 4,73 (m, 1H), 4,65 (m, 1H), 4,6-4,4 (m, 2H), 4,35 (m, 1H, hoved rotamer), 4,21 (dublett av multippelts, 2H), 4,12 (m, 1H, hoved rotamer), 4,06 (m, 1H, mindre rotamer), 3,99 (m, 1H, mindre rotamer), 2,69 (m, 'H-NMR (600 MHz; CD3OD) rotamers: δ 7.8-7.7 (m, 2H), 7.54 (d, 2H), 7.08 (s, 1H, main rotamer), 7.04 (s, 1H, minor rotamer), 6.99 (s, 1H, major rotamer), 6.95 (s, 1H), 6.92 (s, 1H, minor rotamer), 5.18 (m, 1H, minor rotamer), 5.14 (s, 1H, major rotamer), 5.08 (s, 1H, minor rotamer), 4.80 (m, 1H, major rotamer), 4.73 (m, 1H), 4 .65 (m, 1H), 4.6-4.4 (m, 2H), 4.35 (m, 1H, main rotamer), 4.21 (doublet of multiplets, 2H), 4.12 (m, 1H, major rotamer), 4.06 (m, 1H, minor rotamer), 3.99 (m, 1H, minor rotamer), 2.69 (m,

1H, mindre rotamer), 2,53 (m, 1H, hoved rotamer), 2,29 (m, 1H, hoved rotamer), 2,14 (m, 1H, mindre rotamer). 1H, minor rotamer), 2.53 (m, 1H, major rotamer), 2.29 (m, 1H, major rotamer), 2.14 (m, 1H, minor rotamer).

<13>C-NMR (150 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) 8 172,8,172,1, 167,4. <13>C-NMR (150 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms) δ 172.8, 172.1, 167.4.

ESI-MS+: (M+l) = 463 (m/z) ESI-MS+: (M+1) = 463 (m/z)

Eksempel 22 Example 22

Ph( 3- Cl)( 5- OCH7CH7F)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCH7CH7F)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe)

Til en løsning av Ph(3-Cl)(5-OCH2CH2F)-(/?)CH(OH)C(0)OH (818 mg, 3,29 mmol; se Eksempel 21 (v) ovenfor) i DMF (30 ml) under nitrogen ved 0°C ble tilsatt HAze-Pab(OMe)-2HCl (1,43 g, 4,27 mmol), PyBOP (1,89 g, 3,68 mmol) og DIPEA (1,06 g, 8,23 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (15:1) og deretter med EtOAc:EtOH (20:1), hvilket gir tittelforbindelsen (880 mg, 54%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH2CH2F)-(/?)CH(OH)C(0)OH (818 mg, 3.29 mmol; see Example 21 (v) above) in DMF (30 ml) under nitrogen at 0°C was added HAze-Pab(OMe)-2HCl (1.43 g, 4.27 mmol), PyBOP (1.89 g, 3.68 mmol) and DIPEA (1.06 g, 8.23 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl 3 :EtOH (15:1) and then with EtOAc:EtOH (20:1) to give the title compound (880 mg, 54%) as a triturate white foam.

Sm.p.: 65-72°C Melting point: 65-72°C

Rf = 0,60 (10:1 CHCl3:EtOH) Rf = 0.60 (10:1 CHCl3:EtOH)

<]>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,58-7,60 (d, J = 8 Hz, 2H), 7,34 (d, J = 7 Hz, 2H), 7,05-7,08 (m, 2H), 6,95-6,99 (m, 1H), 5,08-5,13 (m, 1H), 4,77-4,82 (m, 1H), 4,60-4,68 (m, 1H), 3,99-4,51 (m, 7H), 3,82 (s, 3H), 2,10-2,75 (m, 2H). <13>C-NMR (150 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) 8 173,3,170,8, 152,5. <]>H NMR (300 MHz, CD3OD, complex mixture of rotamers) 8 7.58-7.60 (d, J = 8 Hz, 2H), 7.34 (d, J = 7 Hz, 2H), 7 .05-7.08 (m, 2H), 6.95-6.99 (m, 1H), 5.08-5.13 (m, 1H), 4.77-4.82 (m, 1H) , 4.60-4.68 (m, 1H), 3.99-4.51 (m, 7H), 3.82 (s, 3H), 2.10-2.75 (m, 2H). <13>C-NMR (150 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms) δ 173.3, 170.8, 152.5.

APCI-MS: (M + 1) = 493 m/z. APCI-MS: (M + 1) = 493 m/z.

Eksempel 23 Example 23

Ph( 3- Cn( 5- OCH( CH7F) 9)-(/ g) CH( OH) C( 0).- Aze- Pab x TFA Ph( 3- Cn( 5- OCH( CH7F) 9)-(/ g) CH( OH) C( 0).- Aze- Pab x TFA

(i) 1. 3- difluorisopropvlmetansulfonat (i) 1. 3-Difluoroisopropylmethanesulfonate

Til en magnetisk omrørt løsning av l,3-difluor-2-propanol (7,0 g, 72,8 mmol) i CH2CI2 (100 ml) under nitrogen ved 0°C ble tilsatt trietylamin (22,1 g, 219 mmol) og metansulfonylklorid (10,0 g, 87,4 mmol). Blandingen ble omrørt ved 0°C i 3 timer. Blandingen ble vasket med 2N HC1 (150 ml) og lagene ble separert. Det vandige laget ble ekstrahert med CH2CI2 (200 ml) og de samlede organiske ekstrakter vasket med saltvann (100 ml), tørket (Na2S04), filtrert og konsentrert 1 vakuum, hvilker gir sub-tittelforbindelsen (11,5 g, 91%) som en gul olje som ble anvendt uten ytterligere rensning. To a magnetically stirred solution of 1,3-difluoro-2-propanol (7.0 g, 72.8 mmol) in CH 2 Cl 2 (100 mL) under nitrogen at 0 °C was added triethylamine (22.1 g, 219 mmol) and methanesulfonyl chloride (10.0 g, 87.4 mmol). The mixture was stirred at 0°C for 3 hours. The mixture was washed with 2N HCl (150 mL) and the layers were separated. The aqueous layer was extracted with CH 2 Cl 2 (200 mL) and the combined organic extracts washed with brine (100 mL), dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (11.5 g, 91%) as a yellow oil which was used without further purification.

<*>H NMR (300 MHz, CDCI3) 5 4,97-5,08 (m, 1H), 4,75-4,77 (m, 2H), 4,59-4,61 (m, 2H), 3,12 (s, 3H). <*>H NMR (300 MHz, CDCl 3 ) δ 4.97-5.08 (m, 1H), 4.75-4.77 (m, 2H), 4.59-4.61 (m, 2H) , 3.12 (p, 3H).

(ii) Ph( 3- Cl)( 5- OCH( CH9F) 7) CHO (ii) Ph(3-Cl)(5-OCH(CH9F)7)CHO

Til en løsning av 3-klor-5-hydroksybenzaldehyd (8,0 g, 50,7 mmol; se Eksempel l(ii) ovenfor) og kaliumkarbonat (9,1 g, 66,0 mmol) i DMF (75 ml) under nitrogen ble det tilsatt en løsning av 1,3-difluorisopropyl metansulfonat (11,5 g, 66,0 mmol; se trinn (i) ovenfor) i DMF (75 ml) dråpevis ved romtemperatur. Blandingen ble oppvarmet til 110°C i 18 timer. Reaksjonsblandingen ble avkjølt til 0°C, hellet i is-kald 2N HC1 (200 ml) og ekstrahert med EtOAc (3 x 250 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum. Den brune oljen ble kromatografert på silikagel under eluering med Heks:EtOAc (4:1), hvilker gir sub-tittelforbindelsen (4,4 g, 37%) som en gul olje. To a solution of 3-chloro-5-hydroxybenzaldehyde (8.0 g, 50.7 mmol; see Example 1(ii) above) and potassium carbonate (9.1 g, 66.0 mmol) in DMF (75 mL) under nitrogen, a solution of 1,3-difluoroisopropyl methanesulfonate (11.5 g, 66.0 mmol; see step (i) above) in DMF (75 mL) was added dropwise at room temperature. The mixture was heated to 110°C for 18 hours. The reaction mixture was cooled to 0°C, poured into ice-cold 2N HCl (200 mL) and extracted with EtOAc (3 x 250 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. The brown oil was chromatographed on silica gel eluting with Hex:EtOAc (4:1) to give the sub-title compound (4.4 g, 37%) as a yellow oil.

'H NMR (300 MHz, CDCI3) 5 9,92 (s, 1H), 7,51 (s, 1H), 7,36 (s, 1H), 7,26 (s, 1H), 4,70-4,89 (m, 3H), 4,63-4,68 (m, 2H). 1 H NMR (300 MHz, CDCl 3 ) δ 9.92 (s, 1H), 7.51 (s, 1H), 7.36 (s, 1H), 7.26 (s, 1H), 4.70- 4.89 (m, 3H), 4.63-4.68 (m, 2H).

(iii) Ph( 3- Cl)( 5- OCH( CH7F) 7)-( R. S) CH( OTMS) CN (iii) Ph( 3- Cl)( 5- OCH( CH7F) 7)-( R. S) CH( OTMS) CN

Til en løsning av Ph(3-Cl)(5-OCH(CH2F)2)CHO (4,4 g, 18,7 mmol; se trinn (ii) ovenfor) og sinkjodid (1,5 g, 4,67 mmol) i CH2CI2 (200 ml) ved 0°C under nitrogen ble det tilsatt trimetylsilylcyanid (3,7 g, 37,3 mmol) dråpevis. Blandingen ble omrørt ved 0°C i 3 timer og natten over ved romtemperatur, deretter fortynnet med H2O (200 ml). Det organiske laget ble separert, tørket (Na2SC>4), filtrert og konsentrert 1 vakuum, hvilker gir sub-tittelforbindelsen (5,5 g, 87%) som en brun olje som ble anvendt uten ytterligere rensning. To a solution of Ph(3-Cl)(5-OCH(CH2F)2)CHO (4.4 g, 18.7 mmol; see step (ii) above) and zinc iodide (1.5 g, 4.67 mmol ) in CH 2 Cl 2 (200 mL) at 0°C under nitrogen was added trimethylsilyl cyanide (3.7 g, 37.3 mmol) dropwise. The mixture was stirred at 0°C for 3 hours and overnight at room temperature, then diluted with H 2 O (200 mL). The organic layer was separated, dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (5.5 g, 87%) as a brown oil which was used without further purification.

<!>H NMR (300 MHz, CDCI3) 5 7,12 (s, 1H), 7,00 (s, 2H), 5,42 (s, 1H), 4,70-4,80 (m, 3H), 4,59-4,64 (m, 2H), 0,26 (s, 9H). <!>H NMR (300 MHz, CDCl 3 ) δ 7.12 (s, 1H), 7.00 (s, 2H), 5.42 (s, 1H), 4.70-4.80 (m, 3H ), 4.59-4.64 (m, 2H), 0.26 (s, 9H).

(iv) Ph( 3- Cl)( 5- OCH( CH2F) 9)-( R. S) CH( OH) C( 0) OH (iv) Ph( 3- Cl)( 5- OCH( CH2F) 9)-( R. S) CH( OH) C( 0) OH

Konsentrert saltsyre (50 ml) ble satt til Ph(3-Cl)(5-OCH(CH2F)2)-(R,S)CH(OTMS)CN (5,5 g, 16,3 mmol; se trinn (iii) ovenfor) og løsningen omrørt ved 100°C i 1,5 timer. Etter avkjøling til romtemperatur, ble reaksjonen ytterligere avkjølt til 0°C, gjort basisk langsomt med 3N NaOH (-200 ml) og vasket med Et20 (3 x 200 ml). Det vandige laget ble surgjort med 2N HC1 (75 ml) og ekstrahert med EtOAc (3 x 200 ml). De samlede EtOAc ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (4,6 g, 100%) som en brun olje som ble anvendt uten ytterligere rensning. Concentrated hydrochloric acid (50 mL) was added to Ph(3-Cl)(5-OCH(CH2F)2)-(R,S)CH(OTMS)CN (5.5 g, 16.3 mmol; see step (iii ) above) and the solution stirred at 100°C for 1.5 hours. After cooling to room temperature, the reaction was further cooled to 0°C, basified slowly with 3N NaOH (-200 mL) and washed with Et 2 O (3 x 200 mL). The aqueous layer was acidified with 2N HCl (75 mL) and extracted with EtOAc (3 x 200 mL). The combined EtOAc extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (4.6 g, 100%) as a brown oil which was used without further purification.

<*>H NMR (300 MHz, CD3OD) 8 7,14 (s, 1H), 7,08 (s, 1H), 7,02 (s, 1H), 5,12 (s, 1H), 4,70-4,90 (m, 3H), 4,52-4,67 (m, 2H). <*>H NMR (300 MHz, CD3OD) δ 7.14 (s, 1H), 7.08 (s, 1H), 7.02 (s, 1H), 5.12 (s, 1H), 4, 70-4.90 (m, 3H), 4.52-4.67 (m, 2H).

(v) Ph( 3- Cn( 5- OCH( CHzF) 7)-( 5) CH( OAc) C( 0) OH ( a) oe Ph( 3- Cl)( 5- OCH( CH9F) ?)-(/ ?) CH( OH) C( Q) OH ( b) (v) Ph( 3- Cn( 5- OCH( CHzF) 7)-( 5) CH( OAc) C( 0) OH ( a) oe Ph( 3- Cl)( 5- OCH( CH9F) ?)- (/ ?) CH( OH) C( Q) OH ( b)

En løsning av Ph(3-Cl)(5-OCH(CH2F)2)-(R,S)CH(OH)C(0)OH (4,6 g, 16,4 mmol; se trinn (iv) ovenfor) og Lipase PS "Amano" (3,0 g) i vinylacetat (150 ml) og MTBE (150 ml) ble oppvarmet ved 70°C under nitrogen i 2,5 d. Reaksjonsblandingen ble avkjølt til romtemperatur, enzymet fjernet ved filtrering gjennom Celite®. Filterkaken ble vasket med EtOAc og filtratet konsentrert i vakuum. Kromatografi på silikagel under eluering med CHCl3:MeOH:Et3N (90:8:2) ga trietylaminsaltet av sub-tittelforbindelse (a) som en gul olje. I tillegg, ble trietylaminsaltet av sub-tittelforbindelse (b) (2,2 g) oppnådd og saltet ble oppløst i H20 (100 ml), surgjort med 2N HC1 og ekstrahert med EtOAc (3 x 200 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (b) (1,4 g, 29%) som en gul olje. A solution of Ph(3-Cl)(5-OCH(CH2F)2)-(R,S)CH(OH)C(0)OH (4.6 g, 16.4 mmol; see step (iv) above ) and Lipase PS "Amano" (3.0 g) in vinyl acetate (150 mL) and MTBE (150 mL) were heated at 70°C under nitrogen for 2.5 d. The reaction mixture was cooled to room temperature, the enzyme removed by filtration through Celite®. The filter cake was washed with EtOAc and the filtrate concentrated in vacuo. Chromatography on silica gel eluting with CHCl3:MeOH:Et3N (90:8:2) gave the triethylamine salt of sub-title compound (a) as a yellow oil. In addition, the triethylamine salt of sub-title compound (b) (2.2 g) was obtained and the salt was dissolved in H 2 O (100 mL), acidified with 2N HCl and extracted with EtOAc (3 x 200 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (b) (1.4 g, 29%) as a yellow oil.

Data for Sub- Tittelforbindelse ( b) : Data for Sub-Title connection (b) :

<*>H NMR (300 MHz, CD3OD) 8 7,14 (s, 1H), 7,08 (s, 1H), 7,02 (s, 1H), 5,12 (s, 1H), 4,70-4,90 (m, 3H), 4,52-4,67 (m, 2H). <*>H NMR (300 MHz, CD3OD) δ 7.14 (s, 1H), 7.08 (s, 1H), 7.02 (s, 1H), 5.12 (s, 1H), 4, 70-4.90 (m, 3H), 4.52-4.67 (m, 2H).

(vi) Ph( 3- Cl)( 5- OCH( CHzF) ?)- r/ e) CH( OH) C( Q)- Aze- Pab( Teoc) (vi) Ph( 3- Cl)( 5- OCH( CHzF) ?)- r/ e) CH( OH) C( Q)- Aze- Pab( Teoc)

Til en løsning av Ph(3-Cl)(5-OCH(CH2F)2)-(/?)CH(OH)C(0)OH (824 mg, 2,94 mmol; se trinn (v) ovenfor) i DMF (30 ml) under nitrogen ved 0°C ble det tilsatt HAze-Pab(Teoc)«HCl (1,71 g, 3,81 mmol), PyBOP (1,68 g, 3,23 mmol) og DIPEA (949 mg, 7,34 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (15:1) og deretter med EtOAc:EtOH (20:1), hvilker gir sub-tittelforbindelsen (720 mg, 38%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH(CH2F)2)-(/?)CH(OH)C(0)OH (824 mg, 2.94 mmol; see step (v) above) in DMF (30 mL) under nitrogen at 0°C was added HAze-Pab(Teoc)"HCl (1.71 g, 3.81 mmol), PyBOP (1.68 g, 3.23 mmol) and DIPEA (949 mg, 7.34 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl3:EtOH (15:1) and then with EtOAc:EtOH (20:1) to give the sub-title compound (720 mg, 38%) as a breakable white foam.

Sm.p.: 78-84°C Melting point: 78-84°C

Rf = 0,62 (10:1 CHCl3:EtOH) Rf = 0.62 (10:1 CHCl3:EtOH)

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,79 (d, / = 8 Hz, 2H), 7,42 (d, 7 = 8 Hz, 2H), 7,00-7,12 (m, 3H), 5,08-5,20 (m, 1H), 3,97-4,80 (m, 12H), 2,10-2,75 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s, 9H). 1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) δ 7.79 (d, / = 8 Hz, 2H), 7.42 (d, 7 = 8 Hz, 2H), 7.00-7.12 (m, 3H), 5.08-5.20 (m, 1H), 3.97-4.80 (m, 12H), 2.10-2.75 (m, 2H), 1.05-1 .11 (m, 2H), 0.08 (s, 9H).

APCI-MS: (M + 1) = 639 m/z. APCI-MS: (M + 1) = 639 m/z.

(vii) Ph( 3- Cl)( 5- OCH( CH9F) 2)-(/ ?) CH( OH) C( 0)- Aze- Pab x TFA (vii) Ph( 3- Cl)( 5- OCH( CH9F) 2)-(/ ?) CH( OH) C( 0)- Aze- Pab x TFA

Ph(3-Cl)(5-OCH(CH2F)2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,129 g, 0,202 mmol; se trinn (vi) ovenfor), ble oppløst i 3 ml TFA og fikk reagere i 20 min. TFA ble inndampet og residuet ble fryse tørket fra vann/acetonitril, hvilket gir 0,123 g (100%) av tittelforbindelsen som dens TFA salt. Ph(3-Cl)(5-OCH(CH2F)2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.129 g, 0.202 mmol; see step (vi) above), was dissolved in 3 ml of TFA and allowed to react for 20 min. The TFA was evaporated and the residue was freeze dried from water/acetonitrile to give 0.123 g (100%) of the title compound as its TFA salt.

'H-NMR (400 MHz; CD3OD) rotamerer: 8 7,8-7,7 (m, 2H), 7,55 (d, 2H), 7,2-7,0 (m, 3H), 5,18 (m, 1H, mindre rotamer), 5,15 (s, 1H, hoved rotamer), 5,08 (s, 1H, mindre rotamer), 4,80 (m, 1H, hoved rotamer delvis obscured by CD3OH topp), 4,75-4,4 (m, 7H), 4,38 (m, 1H, hoved rotamer), 4,15 (m, 1H, hoved rotamer), 4,1-3,9 (m, 2H, 2 signaler fra mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,53 (m, 1H, hoved rotamer), 2,30 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer). C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,9,172,6,172,2,171,7,167,1. 1H-NMR (400 MHz; CD3OD) rotamers: δ 7.8-7.7 (m, 2H), 7.55 (d, 2H), 7.2-7.0 (m, 3H), 5, 18 (m, 1H, minor rotamer), 5.15 (s, 1H, major rotamer), 5.08 (s, 1H, minor rotamer), 4.80 (m, 1H, major rotamer partially obscured by CD3OH peak) , 4.75-4.4 (m, 7H), 4.38 (m, 1H, main rotamer), 4.15 (m, 1H, main rotamer), 4.1-3.9 (m, 2H, 2 signals from minor rotamer), 2.70 (m, 1H, minor rotamer), 2.53 (m, 1H, major rotamer), 2.30 (m, 1H, major rotamer), 2.15 (m, 1H , smaller rotamers). C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 172,9,172,6,172,2,171,7,167,1.

ESI-MS+: (M+l) = 495 (m/z) ESI-MS+: (M+1) = 495 (m/z)

Eksempel 24 Example 24

Ph( 3- Cl)( 5- OCH( CHzF) z)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe) Ph( 3- Cl)( 5- OCH( CHzF) z)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe)

Til en løsning av Ph(3-Cl)(5-OCH(CH2F)2)-(/?)CH(OH)C(0)OH (513 mg, 1,83 mmol; se Eksempel 23 (v) ovenfor) i DMF (30 ml) under nitrogen ved 0°C ble det tilsatt HAze-Pab(OMe)-2HCl (797 mg, 2,38 mmol), PyBOP (1,04 g, 2,01 mmol) og DIPEA (591 mg, 4,57 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert 1 vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCtøEtOH (15:1) og deretter med EtOAc:EtOH (20:1), hvilket gir tittelforbindelsen (370 mg, 39%) som et knusbart hvitt skum. To a solution of Ph(3-Cl)(5-OCH(CH2F)2)-(/?)CH(OH)C(0)OH (513 mg, 1.83 mmol; see Example 23 (v) above) in DMF (30 mL) under nitrogen at 0°C was added HAze-Pab(OMe)-2HCl (797 mg, 2.38 mmol), PyBOP (1.04 g, 2.01 mmol) and DIPEA (591 mg , 4.57 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl₃EtOH (15:1) and then with EtOAc:EtOH (20:1) to give the title compound (370 mg, 39%) as a crumbly white foam .

Sm.p.: 58-63°C Melting point: 58-63°C

Rf = 0,66 (10:1 CHCl3:EtOH) Rf = 0.66 (10:1 CHCl3:EtOH)

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,58-7,60 (d, J = 8 Hz, 2H), 7,34 (d, J = 8 Hz, 2H), 7,00-7,12 (m, 3H), 5,08-5,20 (m, 1H), 4,65-4,82 (m, 3H), 4,28-4,65 (m, 5H), 3,92-4,18 (m, 2H), 3,82 (s, 3H), 2,10-2,75 (m, 2H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) 8 7.58-7.60 (d, J = 8 Hz, 2H), 7.34 (d, J = 8 Hz, 2H), 7 .00-7.12 (m, 3H), 5.08-5.20 (m, 1H), 4.65-4.82 (m, 3H), 4.28-4.65 (m, 5H) , 3.92-4.18 (m, 2H), 3.82 (s, 3H), 2.10-2.75 (m, 2H).

<13>C-NMR (150 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) 8 173,2,170,8, 152,5. <13>C-NMR (150 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms) δ 173.2, 170.8, 152.5.

APCI-MS: (M + 1) = 525 m/z. APCI-MS: (M + 1) = 525 m/z.

Eksempel 25 Example 25

Ph( 3- F)( 5- OCHF?)-(/ g) CH( OH) C( Q)- Aze- Pab x TFA Ph( 3- F)( 5- OCHF?)-(/ g) CH( OH) C( Q)- Aze- Pab x TFA

(i) 1 - brom- 3 - fluor- 5- benzyloks vbenzen (i) 1-bromo-3-fluoro-5-benzyloxybenzene

Natriumhydrid (60% dispersjon i olje, 24,0 g, 0,48 mol) ble tilsatt porsjonsvis til en omrørt løsning av vannfri benzylalkohol (64,5 g, 0,60 mol) i THF (1,0 L). Etter at blandingen ble omrørt i 1 time, ble en løsning av 1 -brom-3,5-difluorbenzen (76,8 g, 0,40 mmol) i THF (100 ml) tilsatt dråpevis over en periode på 1 time. Reaksjonsblandingen ble omrørt ved romtemperatur i 2 d. Vann (400 ml) ble tilsatt og THF ble fjernet i vakuum. Det vandige laget ble ekstrahert med heksan (3 x 150 ml). De samlede organiske ekstrakter ble vasket med 2N NaOH (2 x 100 ml) deretter, tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (110,7 g, 98%) som en lysegul olje som ble anvendt uten ytterligere rensning. Sodium hydride (60% dispersion in oil, 24.0 g, 0.48 mol) was added portionwise to a stirred solution of anhydrous benzyl alcohol (64.5 g, 0.60 mol) in THF (1.0 L). After the mixture was stirred for 1 hour, a solution of 1-bromo-3,5-difluorobenzene (76.8 g, 0.40 mmol) in THF (100 mL) was added dropwise over a period of 1 hour. The reaction mixture was stirred at room temperature for 2 d. Water (400 mL) was added and the THF was removed in vacuo. The aqueous layer was extracted with hexane (3 x 150 mL). The combined organic extracts were washed with 2N NaOH (2 x 100 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (110.7 g, 98%) as a pale yellow oil which was used without further purification.

Rf = 0,47 (Heks) Rf = 0.47 (Hex)

<*>H NMR (300 MHz, CDCI3) 8 7,36-7,41 (m, 5H), 6,94 (bs, 1H), 6,87 (d, <*>H NMR (300 MHz, CDCl 3 ) δ 7.36-7.41 (m, 5H), 6.94 (bs, 1H), 6.87 (d,

7H-f = 8 Hz, 1H), 6,63 (d, /H-f = 10 Hz, 1H), 5,03 (s, 2H). 7H-f = 8 Hz, 1H), 6.63 (d, /H-f = 10 Hz, 1H), 5.03 (s, 2H).

(ii) 3- brom- 5- fluorfenol (ii) 3-bromo-5-fluorophenol

Til en løsning av l-brom-3-fluor-5-benzyloksybenzen (110,0 g, 0,39 mol; se trinn (i) ovenfor) og A^iV-dimetylanilin (474,0 g, 3,92 mol) i vannfri CH2C12 (1,0 L) ved 0°C ble tilsatt aluminiumklorid (156,0 g, 1,17 mol). Etter 10 min, ble isbadet fjernet og omrøringen ble fortsatt i 2 timer. Reaksjonen ble stanset ved forsiktig tilsetning av 3N HC1 (600 ml). Lagene ble separert og det vandige laget ble ekstrahert med CH2CI2 (2 x 150 ml). De samlede organiske ekstrakter ble vasket med 2N HC1 (250 ml) og H2O (3 x 250 ml). Til det organiske laget ble tilsatt 15% KOH (500 ml) og lagene ble separert. Det organiske laget ble videre ekstrahert med 2 N KOH (2 x 70 ml). De samlede vandig lag ble vasket med CH2CI2 (3 x 100 ml) og deretter surgjort med 4N HC1. Det vandige laget ble ekstrahert med Et20 (3 x 125 ml) deretter, ble de samlede Et20 ekstrakter tørket To a solution of 1-bromo-3-fluoro-5-benzyloxybenzene (110.0 g, 0.39 mol; see step (i) above) and N-dimethylaniline (474.0 g, 3.92 mol) in anhydrous CH 2 Cl 2 (1.0 L) at 0°C was added aluminum chloride (156.0 g, 1.17 mol). After 10 min, the ice bath was removed and stirring was continued for 2 h. The reaction was quenched by careful addition of 3N HCl (600 mL). The layers were separated and the aqueous layer was extracted with CH 2 Cl 2 (2 x 150 mL). The combined organic extracts were washed with 2N HCl (250 mL) and H 2 O (3 x 250 mL). To the organic layer was added 15% KOH (500 mL) and the layers were separated. The organic layer was further extracted with 2 N KOH (2 x 70 mL). The combined aqueous layers were washed with CH 2 Cl 2 (3 x 100 mL) and then acidified with 4N HCl. The aqueous layer was extracted with Et 2 O (3 x 125 mL) then the combined Et 2 O extracts were dried

(Na2SC«4), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (69,0 g, 92%) som en brun olje som ble anvendt uten ytterligere rensning. (Na 2 SC 4 ), filtered and concentrated in vacuo to give the sub-title compound (69.0 g, 92%) as a brown oil which was used without further purification.

Sm.p.: 33-35°C Melting point: 33-35°C

Rf = 0,25 (CHC13) Rf = 0.25 (CHCl 3 )

<*>H NMR (300 MHz, DMSO-^j) 8 10,38 (s, 1H), 6,90 (dd, 7H.F = 11 Hz, 7 = 2 Hz, lH), 6,81 (s, 1H), 6,59 (dt, 7H-f = 11 Hz, 7 = 2 Hz, lH). <*>H NMR (300 MHz, DMSO-^j) δ 10.38 (s, 1H), 6.90 (dd, 7H.F = 11 Hz, δ = 2 Hz, 1H), 6.81 (s , 1H), 6.59 (dt, 7H-f = 11 Hz, 7 = 2 Hz, 1H).

APCI-MS: (M-l) = 189 m/z APCI-MS: (M-1) = 189 m/z

(iii) 1 - brom- 3 - lfuor- 5- difluormetoks vbenzen (iii) 1-bromo-3-fluoro-5-difluoromethoxybenzene

En blanding av 3-brom-5-fluorfenol (6,1 g, 31,0 mmol; se trinn (ii) ovenfor) og klordifluormetan (13,0 g, 150,0 mmol) i i-PrOH (100 ml) og 30% KOH (80 ml) ble oppvarmet i en forseglet kolbe i 18 timer ved 80-85°C. Reaksjonsblandingen ble avkjølt til romtemperatur og lagene ble separert. Det organiske laget ble konsentrert i vakuum, hvilket gir en fargeløs olje. Det vandige laget ble ekstrahert med Et20 (3 x 30 ml). Den rå oljen og de samlede organiske ekstrakter ble vasket med 2N NaOH (3 x 30 ml) og H2O (3 x 30 ml). De organiske faser ble deretter tørket (Na2S04), filtrert gjennom en liten silikagel plugg og konsentrert 1" vakuum, hvilker gir sub- tittelforbindelsen (6,1 g, 79%) som en fargeløs olje som ble anvendt uten ytterligere rensning. A mixture of 3-bromo-5-fluorophenol (6.1 g, 31.0 mmol; see step (ii) above) and chlorodifluoromethane (13.0 g, 150.0 mmol) in i -PrOH (100 mL) and 30% KOH (80 mL) was heated in a sealed flask for 18 hours at 80-85°C. The reaction mixture was cooled to room temperature and the layers were separated. The organic layer was concentrated in vacuo to give a colorless oil. The aqueous layer was extracted with Et 2 O (3 x 30 mL). The crude oil and combined organic extracts were washed with 2N NaOH (3 x 30 mL) and H 2 O (3 x 30 mL). The organic phases were then dried (Na 2 SO 4 ), filtered through a small plug of silica gel and concentrated 1" vacuum to give the sub-title compound (6.1 g, 79%) as a colorless oil which was used without further purification.

'H NMR (300 MHz, CDCI3) 8 7,11-7,14 (m, 2H), 6,84 (dt, 7 = 9 Hz, 7 = 2 Hz, 1H), 6,50 (t, 7H-f=72Hz, lH). 1 H NMR (300 MHz, CDCl 3 ) δ 7.11-7.14 (m, 2H), 6.84 (dt, 7 = 9 Hz, 7 = 2 Hz, 1H), 6.50 (t, 7H- f=72Hz, 1H).

(iv) l- fluor- 3- difluormetoksv- 5- vinvlbenzen (iv) 1-fluoro-3-difluoromethoxy-5-vinylbenzene

Tri(butyl)vinylstannan (7,0 g, 22,2 mmol) ble satt til en suspensjon av 1 -brom-3-fluor-5-difluormetoksybenzen (4,9 g, 20,2 mmol; se trinn (iii) ovenfor), Tri(butyl)vinylstannane (7.0 g, 22.2 mmol) was added to a suspension of 1-bromo-3-fluoro-5-difluoromethoxybenzene (4.9 g, 20.2 mmol; see step (iii) above ),

diklorbis(trifenylfosfin)palladium(II) (1,42 g, 2,02 mmol) og vannfri litiumklorid (0,90 g, 20,2 mmol) i THF (40 ml) under nitrogen ved 65°C og blandingen ble omrørt i 5 timer. Reaksjonsblandingen ble avkjølt til 0°C og IN NaOH (90 ml) ble tilsatt. Den bifasiske blandingen ble kraftig omrørt i 1 time og deretter ble lagene separert. Det vandige laget ble ekstrahert med Et20 (3 x 70 ml). De samlede organiske lag ble vasket med 2N NaOH (2 x 40 ml) og H2O (40 ml) deretter tørket (Na2S04), filtrert og konsentrert i vakuum. dichlorobis(triphenylphosphine)palladium(II) (1.42 g, 2.02 mmol) and anhydrous lithium chloride (0.90 g, 20.2 mmol) in THF (40 mL) under nitrogen at 65 °C and the mixture was stirred in 5 hours. The reaction mixture was cooled to 0°C and 1N NaOH (90 mL) was added. The biphasic mixture was stirred vigorously for 1 hour and then the layers were separated. The aqueous layer was extracted with Et 2 O (3 x 70 mL). The combined organic layers were washed with 2N NaOH (2 x 40 mL) and H 2 O (40 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo.

Flash kromatografi på silikagel under eluering med heksan ga sub-tittelforbindelsen (2,2 g, 57%) som en fargeløs olje. Flash chromatography on silica gel eluting with hexane gave the sub-title compound (2.2 g, 57%) as a colorless oil.

Rf = 0,47 (Heks) Rf = 0.47 (Hex)

'H NMR (300 MHz, CDC13) 5 6,93-6,99 (m, 2H), 6,73-6,78 (m, 1H), 6,67 (dd, J = 18 Hz, / = 11 Hz, 1H), 6,51 (t, 7H-f = 73 Hz, 1H), 5,77 (d, J = 18 Hz, 1H), 5,36 (d, J = 11 Hz, 1H). 1 H NMR (300 MHz, CDCl 3 ) δ 6.93-6.99 (m, 2H), 6.73-6.78 (m, 1H), 6.67 (dd, J = 18 Hz, / = 11 Hz, 1H), 6.51 (t, 7H-f = 73 Hz, 1H), 5.77 (d, J = 18 Hz, 1H), 5.36 (d, J = 11 Hz, 1H).

(v) Ph( 3- F)( 5- OCHF7)-(/ g) CH( OH) CH2OH (v) Ph( 3- F)( 5- OCHF7)-(/ g) CH( OH) CH2OH

2-metyl-2-propanol (140 ml), H2O (140 ml) og AD-mix-P (39,2 g) ble samlet sammen og avkjølt til 0°C. l-fluor-3-difluormetoksy-5-vinylbenzen (5,0 g, 26,4 mmol; se trinn (iv) ovenfor) oppløst i en liten mengde av 2-metyl-2-propanol ble tilsatt på én gang og heterogenoppslemningen ble kraftig omrørt ved 0°C inntil TLC avslørte fravær av utgangsmaterialet. Reaksjonen ble stanset ved 0°C ved tilsetning av natriumsulfitt (42,0 g) og deretter oppvarmet til romtemperatur og omrørt i 60 min. Reaksjonsblandingen ble ekstrahert med Et20 (3 x 120 ml). De samlede organiske ekstrakter ble tørket (Na2SC*4), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:EtOAc (3:2) ga sub-tittelforbindelse (5,8 g, 98%) som en fargeløs olje. 2-Methyl-2-propanol (140 mL), H 2 O (140 mL) and AD-mix-P (39.2 g) were combined and cooled to 0°C. 1-Fluoro-3-difluoromethoxy-5-vinylbenzene (5.0 g, 26.4 mmol; see step (iv) above) dissolved in a small amount of 2-methyl-2-propanol was added at once and the heterogeneous slurry was vigorously stirred at 0°C until TLC revealed the absence of the starting material. The reaction was quenched at 0°C by addition of sodium sulphite (42.0 g) and then warmed to room temperature and stirred for 60 min. The reaction mixture was extracted with Et 2 O (3 x 120 mL). The combined organic extracts were dried (Na 2 SC* 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl 3 :EtOAc (3:2) gave the sub-title compound (5.8 g, 98%) as a colorless oil.

Rf = 0,41 (3:2 CHCl3:EtOAc) Rf = 0.41 (3:2 CHCl3:EtOAc)

'H NMR (300 MHz, CDC13) 8 6,96-6,99 (m, 2H), 6,77-6,82 (m, 1H), 6,51 (t, /H-f = 73 Hz, 1H), 4,79-4,85 (m, 1H), 3,76-3,84 (m, 1H), 3,58-3,66 (m, 1H), 2,66 (d, J = 3 Hz, 1H), 2,00 (t,7=6Hz, 1H). 1 H NMR (300 MHz, CDCl 3 ) δ 6.96-6.99 (m, 2H), 6.77-6.82 (m, 1H), 6.51 (t, /H-f = 73 Hz, 1H) , 4.79-4.85 (m, 1H), 3.76-3.84 (m, 1H), 3.58-3.66 (m, 1H), 2.66 (d, J = 3 Hz , 1H), 2.00 (t,7=6Hz, 1H).

HPLC Analyse: 89,2%, >99% ee, ChiralPak AD Kolonne (95:5 Heks:EtOH mobil fase). HPLC Analysis: 89.2%, >99% ee, ChiralPak AD Column (95:5 Hex:EtOH mobile phase).

(vi) Ph( 3- F)( 5- OCHF?>(/ ?) CH( OH) CH2OTBS (vi) Ph( 3- F)( 5- OCHF?>(/ ?) CH( OH) CH2OTBS

En løsning av Ph(3-F)(5-OCHF2)-(/?)CH(OH)CH2OH (5,5 g, 24,7 mmol; se trinn (v) ovenfor), 4-(dimetylamino)pyridin (121 mg, 1,0 mmol) og trietylamin (3,0 g, 29,6 mmol) i vannfri CH2C12 (100 ml) ble avkjølt til 0°C. En 1,0 M løsning av tert-butyldimetylsilylklorid i CH2C12 (26,0 ml, 26,0 mmol) ble tilsatt dråpevis og reaksjonsblandingen fikk oppvarmes til romtemperatur og omrørt natten over. Mettet ammoniumklorid-løsning (60 ml) ble tilsatt og lagene ble separert. Det organiske laget ble vasket med mettet ammoniumklorid-løsning (60 ml) og H20 (2 x 35 ml) deretter tørket (Na2SC<4), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:Heks (3:1) ga sub-tittelforbindelsen (7,9 g, 85%) som en gul olje. A solution of Ph(3-F)(5-OCHF2)-(/?)CH(OH)CH2OH (5.5 g, 24.7 mmol; see step (v) above), 4-(dimethylamino)pyridine ( 121 mg, 1.0 mmol) and triethylamine (3.0 g, 29.6 mmol) in anhydrous CH 2 Cl 2 (100 mL) was cooled to 0 °C. A 1.0 M solution of tert-butyldimethylsilyl chloride in CH 2 Cl 2 (26.0 mL, 26.0 mmol) was added dropwise and the reaction mixture was allowed to warm to room temperature and stir overnight. Saturated ammonium chloride solution (60 mL) was added and the layers were separated. The organic layer was washed with saturated ammonium chloride solution (60 mL) and H 2 O (2 x 35 mL) then dried (Na 2 SC<4), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl 3 :Hex (3:1) gave the sub-title compound (7.9 g, 85%) as a yellow oil.

Rf = 0,47 (3:1 CHCl3:Heks) Rf = 0.47 (3:1 CHCl3:Hex)

'H NMR (300 MHz, CDC13) 8 6,95-6,98 (m, 2H), 6,76-6,79 (m, 1H), 6,51 (t, 7H-f = 73 Hz, 1H), 4,71-4,74 (m, 1H), 3,75-3,80 (m, 1H), 3,48-3,54 (m, 1H), 2,99 (bs, 1H), 0,91 (s, 9H), 0,05 (s, 3H), 0,00 (s, 3H). 1 H NMR (300 MHz, CDCl 3 ) δ 6.95-6.98 (m, 2H), 6.76-6.79 (m, 1H), 6.51 (t, 7H-f = 73 Hz, 1H ), 4.71-4.74 (m, 1H), 3.75-3.80 (m, 1H), 3.48-3.54 (m, 1H), 2.99 (bs, 1H), 0.91 (s, 9H), 0.05 (s, 3H), 0.00 (s, 3H).

(vii) Ph( 3- F)( 5- OCHF?)-(/ ?) CH( OMEM) CH, OTBS (vii) Ph( 3- F)( 5- OCHF?)-(/ ?) CH( OMEM) CH, OTBS

Til en løsning av Ph(3-F)(5-OCHF2)-(/?)CH(OH)CH2OTBS (7,9 g, 0,51 mmol; se trinn (vi) ovenfor) og DIPEA (4,9 g, 48,1 mmol) i vannfri CH2C12 (50 ml) ved 0°C under nitrogen ble det tilsatt dråpevis 2-metoksyetoksymetylklorid (6,6 g, 48,1 mmol). Blandingen ble omrørt i 24 timer. Mettet ammoniumklorid-løsning (70 ml) ble tilsatt og lagene ble separert. Det organiske laget ble vasket med mettet ammoniumklorid-løsning (70 ml) og H20 (3 x 60 ml) deretter tørket (Na2SC»4), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (8,8 g, 99%) som en gul olje som ble anvendt uten ytterligere rensning. To a solution of Ph(3-F)(5-OCHF2)-(/?)CH(OH)CH2OTBS (7.9 g, 0.51 mmol; see step (vi) above) and DIPEA (4.9 g , 48.1 mmol) in anhydrous CH 2 Cl 2 (50 mL) at 0°C under nitrogen was added dropwise 2-methoxyethoxymethyl chloride (6.6 g, 48.1 mmol). The mixture was stirred for 24 hours. Saturated ammonium chloride solution (70 mL) was added and the layers were separated. The organic layer was washed with saturated ammonium chloride solution (70 mL) and H 2 O (3 x 60 mL) then dried (Na 2 SC 4 ), filtered and concentrated in vacuo to give the sub-title compound (8.8 g, 99%) as a yellow oil which was used without further purification.

Rf = 0,41 (4:1 CHCl3:EtOAc) Rf = 0.41 (4:1 CHCl3:EtOAc)

<J>H NMR (300 MHz, CDC13) 8 7,20 (s, 1H), 7,06 (s, 1H), 7,02 (s, 1H), 6,50 (t, 7H-f = 73 Hz, 1H), 4,79-4,81 (m, 1H), 4,66-4,68 (m, 2H), 3,47-3,82 (m, 6H), 3,36 (s, 3H), 0,85 (s, 9H), 0,01 (s, 3H), 0,00 (s, 3H). <J>H NMR (300 MHz, CDCl 3 ) δ 7.20 (s, 1H), 7.06 (s, 1H), 7.02 (s, 1H), 6.50 (t, 7H-f = 73 Hz, 1H), 4.79-4.81 (m, 1H), 4.66-4.68 (m, 2H), 3.47-3.82 (m, 6H), 3.36 (s, 3H), 0.85 (s, 9H), 0.01 (s, 3H), 0.00 (s, 3H).

(viii) Ph( 3- F)( 5- OCHF?)-(/ g) CH( OMEM) CH7QH (viii) Ph( 3- F)( 5- OCHF?)-(/ g) CH( OMEM) CH7QH

Til en løsning av Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)CH2OTBS (9,3 g, 21,9 mmol; se trinn (vii) ovenfor) i THF (60 ml) ved romtemperatur ble det tilsatt en 1,0 M løsning av tetrabutylammonium fluorid i THF (70,0 ml, 70,0 mmol) og blandingen ble omrørt natten over under nitrogen. Reaksjonen ble konsentrert i vakuum. Det gule residuet ble oppløst i Et20 (100 ml) og heksan (100 ml) og vasket suksessivt med mettet ammoniumklorid-løsning (2 x 150 ml) og H20 (3 x 70 ml). Det organiske laget ble tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med Heks:EtOAc (1:1) ga sub-tittelforbindelse (4,2 g, 62%) som en gul olje. To a solution of Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)CH2OTBS (9.3 g, 21.9 mmol; see step (vii) above) in THF (60 mL) at to room temperature, a 1.0 M solution of tetrabutylammonium fluoride in THF (70.0 mL, 70.0 mmol) was added and the mixture was stirred overnight under nitrogen. The reaction was concentrated in vacuo. The yellow residue was dissolved in Et 2 O (100 mL) and hexane (100 mL) and washed successively with saturated ammonium chloride solution (2 x 150 mL) and H 2 O (3 x 70 mL). The organic layer was dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (1:1) gave the sub-title compound (4.2 g, 62%) as a yellow oil.

Rf = 0,42 (1:1 Heks:EtOAc) Rf = 0.42 (1:1 Hex:EtOAc)

'H NMR (300 MHz, CDC13) 5 6,91-6,95 (m, 2H), 6,75-6,81 (m, 1H), 6,51 (t, 7H-f = 73 Hz, 1H), 4,80-4,82 (m, 1H), 4,70-4,74 (m, 2H), 3,88-3,93 (m, 1H), 3,67-3,71 (m, 3H), 3,53-3,56 (m, 2H), 3,39 (s, 3H), 2,96-2,99 (m, 1H). 1 H NMR (300 MHz, CDCl 3 ) δ 6.91-6.95 (m, 2H), 6.75-6.81 (m, 1H), 6.51 (t, 7H-f = 73 Hz, 1H ), 4.80-4.82 (m, 1H), 4.70-4.74 (m, 2H), 3.88-3.93 (m, 1H), 3.67-3.71 (m , 3H), 3.53-3.56 (m, 2H), 3.39 (s, 3H), 2.96-2.99 (m, 1H).

(ix) Ph( 3- F)( 5- OCHF?)-(/ ?) CH( OMEM) C( Q) OH (ix) Ph( 3- F)( 5- OCHF?)-(/ ?) CH( OMEM) C( Q) OH

En løsning av Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)CH2OH (4,2 g, 13,4 mmol; se trinn (viii) ovenfor) i aceton (100 ml) ble satt til en vandig 5% NaHC03 løsning (35 ml). Denne magnetisk omrørte heterogenblandingen ble avkjølt til 0°C og kaliumbromid (159 mg, 1,3 mmol) og 2,2,6,6-tetrametyl-l-piperidinyloksy, fri rest (2,2 g, 14,1 mmol) ble tilsatt. Natriumhypokloritt (5,25%, 30 ml) ble deretter tilsatt dråpevis over en periode på 20 min mens blandingen ble kraftig omrørt og holdt ved 0°C. Etter 1 time, ble rligere natriumhypokloritt (30 ml) og 5% NaHCC>3 løsning (35 ml) tilsatt og omrøringen ble fortsatt ved 0°C i 2 timer. Aceton ble fjernet 1 vakuum. Det vandige laget ble vasket med Et20 (4 x 40 ml). Det vandige laget ble surgjort til pH 3,5 med 10% sitronsyre og ekstrahert med EtOAc (4 x 50 ml). De samlede EtOAc ekstrakter ble suksessivt vasket med H20 (4 x 30 ml) og saltvann (60 ml) deretter, tørket (Na2S04), filtrert og konsentrert 1 vakuum, hvilker gir sub-tittelforbindelsen (4,3 g, 98%) som en fargeløs olje som ble anvendt uten ytterligere rensning. A solution of Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)CH2OH (4.2 g, 13.4 mmol; see step (viii) above) in acetone (100 mL) was added to an aqueous 5% NaHCO 3 solution (35 mL). This magnetically stirred heterogeneous mixture was cooled to 0°C and potassium bromide (159 mg, 1.3 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy, free residue (2.2 g, 14.1 mmol) were added. Sodium hypochlorite (5.25%, 30 mL) was then added dropwise over a period of 20 min while the mixture was vigorously stirred and kept at 0°C. After 1 hour, more sodium hypochlorite (30 mL) and 5% NaHCO3 solution (35 mL) were added and stirring was continued at 0°C for 2 hours. Acetone was removed 1 vacuum. The aqueous layer was washed with Et 2 O (4 x 40 mL). The aqueous layer was acidified to pH 3.5 with 10% citric acid and extracted with EtOAc (4 x 50 mL). The combined EtOAc extracts were successively washed with H 2 O (4 x 30 mL) and brine (60 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (4.3 g, 98%) as a colorless oil which was used without further purification.

Rf = 0,74 (8,0:1,5:0,5 CHCl3:MeOH:Et3N) Rf = 0.74 (8.0:1.5:0.5 CHCl3:MeOH:Et3N)

'H NMR (300 MHz, aceton-^j) 8 7,16-7,18 (m, 2H), 7,16 (t, 7H-f = 89 Hz, 1H), 7,00-7,03 (m, 1H), 5,30 (s, 1H), 4,88 (d, J = 7 Hz, 1H), 4,80 (d, / = 7 Hz, 1H), 3,54-3,75 (m, 2H), 3,46-3,49 (m, 2H), 3,28 (s, 3H). 1 H NMR (300 MHz, acetone-^j) δ 7.16-7.18 (m, 2H), 7.16 (t, 7H-f = 89 Hz, 1H), 7.00-7.03 ( m, 1H), 5.30 (s, 1H), 4.88 (d, J = 7 Hz, 1H), 4.80 (d, / = 7 Hz, 1H), 3.54-3.75 ( m, 2H), 3.46-3.49 (m, 2H), 3.28 (s, 3H).

(x) Ph( 3- F)( 5- OCHF?U/ ftCH( OMEM) C( Q)- Aze- Pab( Teoc) (x) Ph( 3- F)( 5- OCHF?U/ ftCH( OMEM) C( Q)- Aze- Pab( Teoc)

Til en løsning av Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (1,1 g, 3,4 mmol; se trinn (ix) ovenfor) i DMF (20 ml) under nitrogen ved 0°C ble det tilsatt HAze-Pab(Teoc)«HCl (2,0 g, 4,4 mmol), PyBOP (1,9 g, 3,7 mmol) og DIPEA (1,1 g, 8,4 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (15:1) og deretter med EtOAc:EtOH (20:1), hvilker gir sub-tittelforbindelsen (1,3 g, 56%) som et knusbart hvitt skum. To a solution of Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (1.1 g, 3.4 mmol; see step (ix) above) in DMF ( 20 ml) under nitrogen at 0°C was added HAze-Pab(Teoc)"HCl (2.0 g, 4.4 mmol), PyBOP (1.9 g, 3.7 mmol) and DIPEA (1.1 g, 8.4 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl3:EtOH (15:1) and then with EtOAc:EtOH (20:1) to give the sub-title compound (1.3 g, 56% ) as a crushable white foam.

Rf = 0,65 (15:1 CHCl3:EtOH) Rf = 0.65 (15:1 CHCl3:EtOH)

<:>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,80-7,84 (m, 2H), 7,40-7,46 (m, 2H), 6,95-7,16 (m, 3H), 6,92 og 6,88 (t, 7H-f = 73 Hz, 1H), 5,28 og 5,08 (s, 1H), 5,18-5,22 og 4,70-4,78 (m, 1H), 4,50-4,75 (m, 1H), 4,30-4,49 (m, 2H), 4,21-4,26 (m, 3H), 3,97-4,08 (m, 1H), 3,35-3,72 (m, 6H), 3,30 (s, 3H), 2,10-2,75 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s, 9H). <:>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.80-7.84 (m, 2H), 7.40-7.46 (m, 2H), 6.95-7.16 (m, 3H), 6.92 and 6.88 (t, 7H-f = 73 Hz, 1H), 5.28 and 5.08 (s, 1H), 5.18-5.22 and 4.70 -4.78 (m, 1H), 4.50-4.75 (m, 1H), 4.30-4.49 (m, 2H), 4.21-4.26 (m, 3H), 3 .97-4.08 (m, 1H), 3.35-3.72 (m, 6H), 3.30 (s, 3H), 2.10-2.75 (m, 2H), 1.05 -1.11 (m, 2H), 0.08 (s, 9H).

(xi) Ph( 3- F)( 5- OCHF9)-(/ ?) CH( OH) C( 0)- Aze- Pab( Teoc) (xi) Ph( 3- F)( 5- OCHF9)-(/ ?) CH( OH) C( 0)- Aze- Pab( Teoc)

En blanding av Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(Teoc) (590 mg, 0,87 mmol; se trinn (x) ovenfor) og karbontetrabromid (287 mg, 0,87 mmol) i 2-propanol (20 ml) ble tilbakeløpskokt i 1,5 timer. Blandingen ble konsentrert i vakuum deretter, fordelt med H20 (50 ml) og EtOAc (3 x 50 ml). Det vandige laget ble ekstrahert med ytterligere EtOAc (2 x 10 ml). De samlede organiske ekstrakter ble vasket med saltvann (30 ml) deretter tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:EtOH (15:1) ga sub-tittelforbindelsen (60 mg, 12%) som et knusbart hvitt skum. A mixture of Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(Teoc) (590 mg, 0.87 mmol; see step (x) above) and carbon tetrabromide (287 mg, 0.87 mmol) in 2-propanol (20 mL) was refluxed for 1.5 h. The mixture was concentrated in vacuo then partitioned with H 2 O (50 mL) and EtOAc (3 x 50 mL). The aqueous layer was extracted with additional EtOAc (2 x 10 mL). The combined organic extracts were washed with brine (30 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl 3 :EtOH (15:1) afforded the sub-title compound (60 mg, 12%) as a crushable white foam.

Rf = 0,46 (15:1 CHCl3:EtOH) Rf = 0.46 (15:1 CHCl3:EtOH)

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,74 (d, J = 8 Hz, 2H), 7,35-7,37 (m, 2H), 6,97-7,07 (m, 2H), 6,80-6,84 (m, 1H), 6,82 og 6,80 (t, 7H-f = 73 Hz, 1H), 5,10 og 5,06 (s, 1H), 4,68-4,70 (m, 1H), 3,97-4,60 (m, 6H), 2,10-2,75 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s, 9H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.74 (d, J = 8 Hz, 2H), 7.35-7.37 (m, 2H), 6.97-7, 07 (m, 2H), 6.80-6.84 (m, 1H), 6.82 and 6.80 (t, 7H-f = 73 Hz, 1H), 5.10 and 5.06 (s, 1H), 4.68-4.70 (m, 1H), 3.97-4.60 (m, 6H), 2.10-2.75 (m, 2H), 1.05-1.11 ( m, 2H), 0.08 (s, 9H).

APCI-MS: (M + 1) = 595 m/z APCI-MS: (M + 1) = 595 m/z

(xii) Ph( 3- F)( 5- OCHF?)-(/ g) CH( OH) C( Q)- Aze- Pab x TFA (xii) Ph( 3- F)( 5- OCHF?)-(/ g) CH( OH) C( Q)- Aze- Pab x TFA

Ph(3-F)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0,053 g, 0,089 mmol; se trinn (xi) ovenfor), ble oppløst i 3 ml TFA og fikk reagere i 80 min mens den ble avkjølt på et isbad. TFA ble inndampet og residuet ble fryse tørket fra vann/acetonitril, hvilket gir 0,042 g (80%) av tittelforbindelsen som dens TFA salt. Ph(3-F)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (0.053 g, 0.089 mmol; see step (xi) above), was dissolved in 3 ml TFA and allowed to react for 80 min while cooling in an ice bath. The TFA was evaporated and the residue was freeze dried from water/acetonitrile to give 0.042 g (80%) of the title compound as its TFA salt.

'H-NMR (300 MHz; CD3OD) rotamerer: 8 7,7-7,6 (m, 2H), 7,5-7,4 (m, 2H), 7,1-6,6 (m, 4H), 5,2-5,0 (m, 1H pluss mindre rotamer av 1H), ca 4,8 (hoved rotamer av tidligere signal uklart p.g.a.CD3OH signal), 4,6-4,3 (m, 2H), 4,26 (m, 1H, hoved rotamer), 4,10 (m, 1H, hoved rotamer), 3,96 (m, 1H, mindre rotamer), 3,89 (m, 1H, mindre rotamer), 2,60 (m, 1H, mindre rotamer), 2,44 (m, 1H, hoved rotamer), 2,19 (m, 1H, hoved rotamer), 2,05 (m, 1H, mindre rotamer). 1H-NMR (300 MHz; CD 3 OD) rotamers: δ 7.7-7.6 (m, 2H), 7.5-7.4 (m, 2H), 7.1-6.6 (m, 4H ), 5.2-5.0 (m, 1H plus minor rotamer of 1H), approx. 4.8 (main rotamer of previous signal unclear due to CD3OH signal), 4.6-4.3 (m, 2H), 4 .26 (m, 1H, major rotamer), 4.10 (m, 1H, major rotamer), 3.96 (m, 1H, minor rotamer), 3.89 (m, 1H, minor rotamer), 2.60 (m, 1H, minor rotamer), 2.44 (m, 1H, major rotamer), 2.19 (m, 1H, major rotamer), 2.05 (m, 1H, minor rotamer).

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) 8 172,8,172,0, 167,0. <13>C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms) δ 172.8, 172.0, 167.0.

ESI-MS+: (M+l) = 451 (m/z) ESI-MS+: (M+1) = 451 (m/z)

Eksempel 26 Example 26

Ph( 3- F)( 5- OCHF7)-(/ ?) CH( OH) C( Q)- Aze- Pab( OMe) Ph( 3- F)( 5- OCHF7)-(/ ?) CH( OH) C( Q)- Aze- Pab( OMe)

(i) Ph( 3- F)( 5- OCHFz)-(/ ?) CH( OMEM) C( Q)- Aze- Pab( OMe) (i) Ph( 3- F)( 5- OCHFz)-(/ ?) CH( OMEM) C( Q)- Aze- Pab( OMe)

Til en løsning av Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (1,0 g, 3,1 mmol; se Eksempel 25(ix) ovenfor) i DMF (30 ml) under nitrogen ved 0°C ble det tilsatt HAze-Pab(OMe)-2HCl (1,4 g, 4,1 mmol), PyBOP (1,8 g, 3,4 mmol) og DIPEA (1,0 g, 7,8 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (15:1) og deretter med EtOAc, hvilker gir sub-tittelforbindelsen (1,5 g, 79%) som et knusbart hvitt skum. To a solution of Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (1.0 g, 3.1 mmol; see Example 25(ix) above) in DMF (30 mL) under nitrogen at 0°C was added HAze-Pab(OMe)-2HCl (1.4 g, 4.1 mmol), PyBOP (1.8 g, 3.4 mmol) and DIPEA (1, 0 g, 7.8 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl 3 :EtOH (15:1) and then with EtOAc to give the sub-title compound (1.5 g, 79%) as a crumbly white foam.

Rf = 0,24 (EtOAc) Rf = 0.24 (EtOAc)

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,58-7,62 (m, 2H), 7,32-7,38 (m, 2H), 7,03-7,16 (m, 3H), 6,92 og 6,88 (d, 7H-f = 73 Hz, 1H), 5,27 og 5,08 (s, 1H), 5,22-5,15 og 4,75-4,80 (m, 1H), 4,38-4,65 (m, 5H), 3,92-4,27 (m, 1H), 3,82 (s, 3H), 3,43-3,68 (m, 4H), 3,29 (s, 3H), 2,28-2,85 (m, 2H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.58-7.62 (m, 2H), 7.32-7.38 (m, 2H), 7.03-7.16 (m, 3H), 6.92 and 6.88 (d, 7H-f = 73 Hz, 1H), 5.27 and 5.08 (s, 1H), 5.22-5.15 and 4.75 -4.80 (m, 1H), 4.38-4.65 (m, 5H), 3.92-4.27 (m, 1H), 3.82 (s, 3H), 3.43-3 .68 (m, 4H), 3.29 (s, 3H), 2.28-2.85 (m, 2H).

(ii) Ph( 3- F)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- Pab( OMe) (ii) Ph( 3- F)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- Pab( OMe)

En blanding av Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(OMe) (828 mg, 2,33 mmol; se trinn (i) ovenfor) og karbontetrabromid (525 mg, 2,33 mmol) i 2-propanol (20 ml) ble tilbakeløpskokt i 8 timer og deretter omrørt ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet fordelt med H20 (70 ml) og EtOAc (50 ml). Det vandige laget ble ekstrahert med EtOAc (2 x 25 ml). De samlede organiske ekstrakter ble vasket med saltvann (35 ml) deretter, tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:EtOH (15:1) ga tittelforbindelsen (520 mg, 74%) som et knusbart hvitt skum. A mixture of Ph(3-F)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(OMe) (828 mg, 2.33 mmol; see step (i) above) and carbon tetrabromide (525 mg, 2.33 mmol) in 2-propanol (20 mL) was refluxed for 8 hours and then stirred at room temperature overnight. The mixture was concentrated in vacuo and the residue partitioned between H 2 O (70 mL) and EtOAc (50 mL). The aqueous layer was extracted with EtOAc (2 x 25 mL). The combined organic extracts were washed with brine (35 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl 3 :EtOH (15:1) afforded the title compound (520 mg, 74%) as a crushable white foam.

Sm.p.: 73-81°C Melting point: 73-81°C

Rf = 0,43 (15:1 CHCl3:EtOH) Rf = 0.43 (15:1 CHCl3:EtOH)

<!>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,59 (d, / = 8 Hz, 2H), 7,32-7,37 (m, 2H), 7,05-7,14 (m, 2H), 6,87-6,92 (m, 1H), 6,90 og 6,86 (t, 7H-f = 73 Hz, 1H), 5,13-5,18 og 4,75-4,85 (m, 2H), 4,15-4,45 (m, 4H), 3,81 (s, 3H), 2,10-2,75 (m, 2H). <13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) 8 172,0,171,4, 153,9. <!>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.59 (d, / = 8 Hz, 2H), 7.32-7.37 (m, 2H), 7.05-7, 14 (m, 2H), 6.87-6.92 (m, 1H), 6.90 and 6.86 (t, 7H-f = 73 Hz, 1H), 5.13-5.18 and 4, 75-4.85 (m, 2H), 4.15-4.45 (m, 4H), 3.81 (s, 3H), 2.10-2.75 (m, 2H). <13>C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms) δ 172.0, 171.4, 153.9.

APCI-MS: (M + 1) = 481 m/z APCI-MS: (M + 1) = 481 m/z

Eksempel 27 Example 27

Ph( 3- Br)( 5- OCH9F)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA Ph( 3- Br)( 5- OCH9F)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA

(i) 1. 3- dibrom- 5- benzvloksvbenzen (i) 1. 3- dibromo- 5- benzvloxvbenzene

Natriumhydrid (9,9 g, 0,414 mol, 95% tørr) ble tilsatt i porsjoner til en omrørt løsning av benzylalkohol (41,0 g, 0,394 mol) i THF (1,0 L) ved romtemperatur under en nitrogen-atmosfære og omrørt i 1 time. Til denne løsningen ble det tilsatt dråpevis l,3-dibrom-5-fluorbenzen (100,0 g, 0,394 mol). Etter omrøring natten over, ble blandingen fordelt med H2O (600 ml) og EtOAc (4 x 600 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert 1 vakuum. Flash kromatografi på silikagel under eluering med heksaner ga sub-tittelforbindelsen (101,3 g, 75%) som en gul olje. Sodium hydride (9.9 g, 0.414 mol, 95% dry) was added portionwise to a stirred solution of benzyl alcohol (41.0 g, 0.394 mol) in THF (1.0 L) at room temperature under a nitrogen atmosphere and stirred for 1 hour. To this solution was added dropwise 1,3-dibromo-5-fluorobenzene (100.0 g, 0.394 mol). After stirring overnight, the mixture was partitioned with H 2 O (600 mL) and EtOAc (4 x 600 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes afforded the sub-title compound (101.3 g, 75%) as a yellow oil.

'H NMR (300 MHz, CDC13) 8 7,30-7,48 (m, 5H), 7,18 (s, 1H), 7,06 (s, 2H), 4,99 (s, 2H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.30-7.48 (m, 5H), 7.18 (s, 1H), 7.06 (s, 2H), 4.99 (s, 2H).

(ii) 3. 5- dibromfenol (ii) 3. 5- dibromophenol

Aluminiumklorid (11,7 g, 87,6 mmol) ble tilsatt i porsjoner til en løsning av l,3-dibrom-5-benzyloksybenzen (10,0 g, 29,2 mmol; se trinn (i) ovenfor) og iV^V-dimetylanilin (35,4 g, 292 mmol) i CH2CI2 (100 ml) ved romtemperatur under en nitrogen-atmosfære. Etter 30 min, ble blandingen fordelt med IN HC1 (300 ml) og EtOAc (5 x 150 ml). De samlede organiske ekstrakter ble vasket med mettet NaHC03 (150 ml) og saltvann (150 ml) deretter, tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med Heks:EtOAc (9:1) ga sub-tittelforbindelsen (6,1 g, 82%) som et hvitt, fast stoff. Aluminum chloride (11.7 g, 87.6 mmol) was added portionwise to a solution of 1,3-dibromo-5-benzyloxybenzene (10.0 g, 29.2 mmol; see step (i) above) and iV^ V-dimethylaniline (35.4 g, 292 mmol) in CH 2 Cl 2 (100 mL) at room temperature under a nitrogen atmosphere. After 30 min, the mixture was partitioned with 1N HCl (300 mL) and EtOAc (5 x 150 mL). The combined organic extracts were washed with saturated NaHCO 3 (150 mL) and brine (150 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (9:1) gave the sub-title compound (6.1 g, 82%) as a white solid.

'H NMR (300 MHz, CDC13) 8 7,21 (s, 1H), 6,97 (s, 2H), 5,88 (bs, 1H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.21 (s, 1H), 6.97 (s, 2H), 5.88 (bs, 1H).

(iii) 1. 3- dibrom- 5- monolfuormetoksvbenzen (iii) 1. 3- dibromo- 5- monolfluoromethoxybenzene

Til en tarert, forseglet 350 ml rundbunnet trykk kolbe inneholdende en suspensjon av 3,5-dibromfenol (10,0 g, 39,7 mmol; se trinn (ii) ovenfor) og CS2CO3 (20,7 g, 63,5 mmol) i DMF (150 ml)ved -78°C ble det tilsatt klorfluormetan via bobling i 5 min gjennom septum. Septumet ble erstattet med en Teflon stopper og kolben ble deretter forseglet og fikk oppvarmes til romtemperatur hvor kolben ble veiet og bestemt å inneholde 9,0 g (131 mmol) klorfluormetan. Løsningen ble oppvarmet i et oljebad satt til 70°C natten over. Kolben ble avkjølt til romtemperatur, trykket forsiktig frigjort og innholdet fortynnet med vann (100 ml). Det vandige laget ble ekstrahert med Et20 (3 x 200 ml) deretter, de samlede organiske lag ble tørket (Na2SC>4), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med heksaner ga sub-tittelforbindelse (7,9 g, 71%) som et hvitt, fast stoff. To a tared, sealed 350 mL round-bottomed pressure flask containing a suspension of 3,5-dibromophenol (10.0 g, 39.7 mmol; see step (ii) above) and CS2CO3 (20.7 g, 63.5 mmol) in DMF (150 ml) at -78°C was added chlorofluoromethane via bubbling for 5 min through the septum. The septum was replaced with a Teflon stopper and the flask was then sealed and allowed to warm to room temperature where the flask was weighed and determined to contain 9.0 g (131 mmol) of chlorofluoromethane. The solution was heated in an oil bath set at 70°C overnight. The flask was cooled to room temperature, the pressure carefully released and the contents diluted with water (100 mL). The aqueous layer was extracted with Et 2 O (3 x 200 mL) then the combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes gave the sub-title compound (7.9 g, 71%) as a white solid.

'H NMR (300 MHz, CDCI3) 5 7,40 (s, 1H), 7,18 (s, 2H), 5,67 (d, 7H-f = 53 Hz, 2H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.40 (s, 1H), 7.18 (s, 2H), 5.67 (d, 7H-f = 53 Hz, 2H).

(iv) 1 - brom- 3 - monofluormetoks v- 5- vinvlbenzen (iv) 1-bromo-3-monofluoromethoxy v-5-vinylbenzene

Tri(butyl)vinyltin (10,0 g, 31,4 mmol) ble satt dråpevis til en løsning av l,3-dibrom-5-monofluormetoksybenzen (8,5 g, 29,9 mmol; se trinn (iii) ovenfor), tetrakis(trifenylfosfin)palladium(0) (690 mg, 0,599 mmol) og 2,6-di-ferf-butyl-4-metylfenol (spatel tip) i toluen (100 ml) under nitrogen. Blandingen ble omrørt ved 70°C i 8 timer. Blandingen ble avkjølt til 0°C og IN NaOH (70 ml) ble tilsatt. Etter 1 time, ble blandingen ekstrahert med CH2CI2 (3 x 300 ml) deretter, ble de samlede organiske lag tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med heksaner ga sub-tittelforbindelsen (4,3 g, 57%) som en fargeløs olje. Tri(butyl)vinyltin (10.0 g, 31.4 mmol) was added dropwise to a solution of 1,3-dibromo-5-monofluoromethoxybenzene (8.5 g, 29.9 mmol; see step (iii) above) , tetrakis(triphenylphosphine)palladium(0) (690 mg, 0.599 mmol) and 2,6-di-tert-butyl-4-methylphenol (spatula tip) in toluene (100 mL) under nitrogen. The mixture was stirred at 70°C for 8 hours. The mixture was cooled to 0°C and 1N NaOH (70 mL) was added. After 1 hour, the mixture was extracted with CH 2 Cl 2 (3 x 300 mL) then the combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes afforded the sub-title compound (4.3 g, 57%) as a colorless oil.

'H NMR (300 MHz, CDC13) 8 7,30 (s, 1H), 7,16 (s, 1H), 7,01 (s, 1H), 6,60 (dd, 7 = 6 Hz, 7 = 11 Hz, 1H), 5,74 (d, 7 = 16 Hz, 1H), 5,67 (d, 7H-f = 53 Hz, 2H), 5,32 (d, 7 = 8 Hz, 1H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.30 (s, 1H), 7.16 (s, 1H), 7.01 (s, 1H), 6.60 (dd, 7 = 6 Hz, 7 = 11 Hz, 1H), 5.74 (d, 7 = 16 Hz, 1H), 5.67 (d, 7H-f = 53 Hz, 2H), 5.32 (d, 7 = 8 Hz, 1H).

(v) Ph( 3- Br)( 5- OCH9F)-(/ g) CH( OH) CH7QH (v) Ph( 3- Br)( 5- OCH9F)-(/ g) CH( OH) CH7QH

2-metyl-2-propanol (100 ml), H20 (100 ml) og AD-mix-Ø (27,5 g) ble samlet sammen og avkjølt til 0°C. l-brom-3-monofluormetoksy-5-vinylbenzen (4,3 g, 17,3 mmol; se trinn (iv) ovenfor) ble tilsatt på én gang og heterogenoppslemningen ble kraftig omrørt ved 0°C inntil TLC avslørte fravær av utgangsmaterialet. Reaksjonen ble stanset ved 0°C ved tilsetning av mettet natriumsulfitt (200 ml) og deretter oppvarmet til romtemperatur og 2-Methyl-2-propanol (100 mL), H 2 O (100 mL) and AD-mix-Ø (27.5 g) were combined and cooled to 0°C. 1-Bromo-3-monofluoromethoxy-5-vinylbenzene (4.3 g, 17.3 mmol; see step (iv) above) was added in one portion and the heterogeneous slurry was vigorously stirred at 0°C until TLC revealed the absence of the starting material. The reaction was quenched at 0°C by the addition of saturated sodium sulfite (200 mL) and then warmed to room temperature and

omrørt i 60 min. Reaksjonsblandingen ble ekstrahert med EtOAc (3 x 150 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (4,9 g, 100%) som en fargeløs olje som ble anvendt uten ytterligere rensning. stirred for 60 min. The reaction mixture was extracted with EtOAc (3 x 150 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (4.9 g, 100%) as a colorless oil which was used without further purification.

'H NMR (300 MHz, CD3OD) 8 7,30 (s, 1H), 7,15 (s, 1H), 7,11 (s, 1H), 5,70 (d, JH. F = 53 Hz, 2H), 4,62-4,70 (m, 1H), 3,52-3,70 (m, 2H). 1 H NMR (300 MHz, CD 3 OD) δ 7.30 (s, 1H), 7.15 (s, 1H), 7.11 (s, 1H), 5.70 (d, JH. F = 53 Hz, 2H), 4.62-4.70 (m, 1H), 3.52-3.70 (m, 2H).

HPLC Analyse: 92,1%, 96,9% ee, ChiralPak AD Kolonne (95:5 Heks:EtOH mobil fase). HPLC Analysis: 92.1%, 96.9% ee, ChiralPak AD Column (95:5 Hex:EtOH mobile phase).

(vi) Ph( 3- Br)( 5- OCHzF)-(/ e) CH( OMEM) CHzOTBS (vi) Ph( 3- Br)( 5- OCHzF)-(/ e) CH( OMEM) CHzOTBS

Til en løsning av Ph(3-Br)(5-OCH2F)-(Æ)CH(OH)CH2OH (4,9 g, 18,6 mmol; se trinn (v) ovenfor), 4-(dimetylamino)pyridin (453 mg, 3,71 mmol) og DIPEA (8,9 g, 93,0 mmol) i vannfri CH2C12 (200 ml) ble det tilsatt dråpevis en 1,0 M løsning av tert-butyldimetylsilylklorid i CH2C12 (22,3 ml, 22,3 mmol). Reaksjonsblandingen ble omrørt 10 timer ved romtemperatur. Til blandingen ble det tilsatt DIPEA (8,9 g, 93,0 mmol) og 2-metoksyetoksymetylklorid (13,9 g, 111 mmol) dråpevis. Etter 16 timer, ble ytterligere 2-metoksyetoksymetylklorid (2,2 g) tilsatt og reaksjonsblandingen omrørt natten over. Blandingen ble fortynnet med H20 (100 ml) og lagene ble separert. Det vandige laget ble ekstrahert med CH2C12 (3 x 200 ml) deretter, ble de samlede organiske lag tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med Heks:EtOAc (5:1) ga sub-tittelforbindelsen (4,8 g, 55%) som en fargeløs olje. To a solution of Ph(3-Br)(5-OCH2F)-(Æ)CH(OH)CH2OH (4.9 g, 18.6 mmol; see step (v) above), 4-(dimethylamino)pyridine ( 453 mg, 3.71 mmol) and DIPEA (8.9 g, 93.0 mmol) in anhydrous CH 2 Cl 2 (200 mL) was added dropwise a 1.0 M solution of tert-butyldimethylsilyl chloride in CH 2 Cl 2 (22.3 mL, 22.3 mmol). The reaction mixture was stirred for 10 hours at room temperature. To the mixture was added DIPEA (8.9 g, 93.0 mmol) and 2-methoxyethoxymethyl chloride (13.9 g, 111 mmol) dropwise. After 16 hours, additional 2-methoxyethoxymethyl chloride (2.2 g) was added and the reaction mixture was stirred overnight. The mixture was diluted with H 2 O (100 mL) and the layers were separated. The aqueous layer was extracted with CH 2 Cl 2 (3 x 200 mL) then the combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (5:1) gave the sub-title compound (4.8 g, 55%) as a colorless oil.

'H NMR (300 MHz, CDC13) 8 7,29 (s, 1H), 7,22 (s, 1H), 7,05 (s, 1H), 5,74 (d, 7H-f = 53 Hz, 2H), 4,84 (d, J = 7 Hz, 1H), 4,70-4,74 (m, 2H), 3,50-3,91 (m, 6H), 3,42 (s, 3H), 0,90 (s, 9H), 0,05 (s, 3H), 0,01 (s, 3H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.29 (s, 1H), 7.22 (s, 1H), 7.05 (s, 1H), 5.74 (d, 7H-f = 53 Hz, 2H), 4.84 (d, J = 7 Hz, 1H), 4.70-4.74 (m, 2H), 3.50-3.91 (m, 6H), 3.42 (s, 3H ), 0.90 (s, 9H), 0.05 (s, 3H), 0.01 (s, 3H).

(vii) Ph( 3- Br)( 5- OCH7F)-(/ ?) CH( OMEM) CH2OH (vii) Ph( 3- Br)( 5- OCH7F)-(/ ?) CH( OMEM) CH2OH

Til en løsning av Ph(3-Br)(5-OCH2F)-(Æ)CH(OMEM)CH2OTBS (4,7 g, 10,1 mmol; se trinn (vi) ovenfor) i THF (100 ml) ble det tilsatt en 1,0 M løsning av tetrabutylammoniumfluorid i THF (13,1 ml, 13,1 mmol) ved romtemperatur og blandingen ble omrørt i 1 time. Blandingen ble fordelt med H20 (100 ml) og EtOAc (3 x 100 ml) deretter, de samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (3,3 g, 92%) som en fargeløs olje som ble anvendt uten ytterligere rensning. To a solution of Ph(3-Br)(5-OCH2F)-(Æ)CH(OMEM)CH2OTBS (4.7 g, 10.1 mmol; see step (vi) above) in THF (100 mL) was added a 1.0 M solution of tetrabutylammonium fluoride in THF (13.1 mL, 13.1 mmol) at room temperature and the mixture was stirred for 1 h. The mixture was partitioned with H 2 O (100 mL) and EtOAc (3 x 100 mL) then the combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (3.3 g, 92%) as a colorless oil which was used without further purification.

'H NMR (300 MHz, CD3OD) 8 7,22 (s, 1H), 7,14 (s, 1H), 7,03 (s, 1H), 5,71 (d, 7H-f = 53 Hz, 2H), 4,80-4,82 (m, 1H), 4,58-4,66 (m, 2H), 3,71-3,77 (m, 1H), 3,39-3,65 (m, 5H), 3,27 (s, 3H). 1 H NMR (300 MHz, CD 3 OD) δ 7.22 (s, 1H), 7.14 (s, 1H), 7.03 (s, 1H), 5.71 (d, 7H-f = 53 Hz, 2H), 4.80-4.82 (m, 1H), 4.58-4.66 (m, 2H), 3.71-3.77 (m, 1H), 3.39-3.65 ( m, 5H), 3.27 (s, 3H).

(viii) Ph( 3- Br)( 5- OCH2F)-(/ g) CH( OMEM) C( 0) OH (viii) Ph( 3- Br)( 5- OCH2F)-(/ g) CH( OMEM) C( 0) OH

En løsning av Ph(3-Br)(5-OCH2F)-(/?)CH(OMEM)CH2OH (2,1 g, 6,0 mmol; se trinn (vii) ovenfor) i aceton (40 ml) ble satt til en vandig 5% NaHCC>3 løsning (15 ml). Denne magnetisk omrørte heterogene blandingen ble avkjølt til 0°C og kaliumbromid (70 mg, 0,60 mmol) og 2,2,6,6-tetrametyl-l-piperidinyloksy, fri rest (976 mg, 5,8 mmol) ble tilsatt. Natriumhypokloritt (5,25%, 15 ml) ble deretter tilsatt dråpevis over en periode på 10 min mens blandingen ble kraftig omrørt og holdt ved 0°C. Etter 1 time, ble ytterligere natriumhypokloritt (10 ml) og NaHCC«3 løsning (20 ml) tilsatt og omrøringen ble fortsatt ved 0°C i ytterligere 4 timer. Aceton ble fjernet på en rotasjonsinndamper. Det vandige laget ble fortynnet med 10% NaHC03 løsning (30 ml) og ble vasket med Et20 (3 x 20 ml). Det vandige laget ble surgjort til pH 3,5 med 10% sitronsyre og ekstrahert med EtOAc (3 x 40 ml). De samlede EtOAc ekstrakter ble vasket med H20 (3 x 50 ml) og saltvann (50 ml) deretter, tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (1,7 g, 78%) som en fargeløs olje som ble anvendt uten ytterligere rensning. A solution of Ph(3-Br)(5-OCH2F)-(/?)CH(OMEM)CH2OH (2.1 g, 6.0 mmol; see step (vii) above) in acetone (40 mL) was added to an aqueous 5% NaHCC>3 solution (15 mL). This magnetically stirred heterogeneous mixture was cooled to 0°C and potassium bromide (70 mg, 0.60 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy, free residue (976 mg, 5.8 mmol) were added. . Sodium hypochlorite (5.25%, 15 mL) was then added dropwise over a period of 10 min while the mixture was vigorously stirred and kept at 0°C. After 1 hour, additional sodium hypochlorite (10 mL) and NaHCCl 3 solution (20 mL) were added and stirring was continued at 0°C for an additional 4 hours. Acetone was removed on a rotary evaporator. The aqueous layer was diluted with 10% NaHCO 3 solution (30 mL) and was washed with Et 2 O (3 x 20 mL). The aqueous layer was acidified to pH 3.5 with 10% citric acid and extracted with EtOAc (3 x 40 mL). The combined EtOAc extracts were washed with H 2 O (3 x 50 mL) and brine (50 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (1.7 g, 78%) as a colorless oil that was used without further purification.

'H NMR (300 MHz, CD3OD) 8 7,38 (s, 1H), 7,25 (s, 1H), 7,18 (s, 1H), 5,76 (d, 7H-f = 53 Hz, 2H), 5,21 (s, 1H), 4,83 (d, J = 7 Hz, 1H), 4,75 (d, J = 7 Hz, 1H), 3,62-3,78 (m, 2H), 3,48-3,52 (m, 2H), 3,32 (s, 3H). 1 H NMR (300 MHz, CD 3 OD) δ 7.38 (s, 1H), 7.25 (s, 1H), 7.18 (s, 1H), 5.76 (d, 7H-f = 53 Hz, 2H), 5.21 (s, 1H), 4.83 (d, J = 7 Hz, 1H), 4.75 (d, J = 7 Hz, 1H), 3.62-3.78 (m, 2H), 3.48-3.52 (m, 2H), 3.32 (s, 3H).

(ix) Ph( 3- Br)( 5- OCHzF)-(/ g) CH( OMEM) C( 0)- Aze- Pab( Teoc) (ix) Ph( 3- Br)( 5- OCHzF)-(/ g) CH( OMEM) C( 0)- Aze- Pab( Teoc)

Til en løsning av Ph(3-Br)(5-OCH2F)-(Æ)CH(OMEM)C(0)OH (1,0 g, 2,72 mmol; se trinn (viii) ovenfor) i DMF (20 ml) under nitrogen ved 0°C ble det tilsatt HAze-Pab(Teoc)»HCl (1,6 g, 3,5 mmol), PyBOP (1,6 g, 3,0 mmol) og DIPEA (880 mg, 6,81 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCtøEtOH (15:1) og deretter med EtOAc:EtOH (20:1), hvilker gir sub-tittelforbindelsen (1,2 g, 62%) som et knusbart hvitt skum. To a solution of Ph(3-Br)(5-OCH2F)-(Æ)CH(OMEM)C(0)OH (1.0 g, 2.72 mmol; see step (viii) above) in DMF (20 ml) under nitrogen at 0°C was added HAze-Pab(Teoc)»HCl (1.6 g, 3.5 mmol), PyBOP (1.6 g, 3.0 mmol) and DIPEA (880 mg, 6 .81 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl2EtOH (15:1) and then with EtOAc:EtOH (20:1) to give the sub-title compound (1.2 g, 62%) as a breakable white foam.

'H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,80-7,84 (m, 2H), 7,40-7,46 (m, 2H), 7,13-7,32 (m, 3H), 5,84-5,87 (m, 1H), 5,67-5,69 (m, 1H), 5,25 og 5,07 (s, 1H), 5,18-5,23 og 4,80-4,88 (m, 1H), 3,97-4,79 (m, 8H), 3,60-3,71 (m, 2H), 3,40-3,53 (m, 2H), 3,32 (s, 3H), 2,10-2,75 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s, 9H). 1 H NMR (300 MHz, CD 3 OD, complex mixture of rotamers) δ 7.80-7.84 (m, 2H), 7.40-7.46 (m, 2H), 7.13-7.32 (m , 3H), 5.84-5.87 (m, 1H), 5.67-5.69 (m, 1H), 5.25 and 5.07 (s, 1H), 5.18-5.23 and 4.80-4.88 (m, 1H), 3.97-4.79 (m, 8H), 3.60-3.71 (m, 2H), 3.40-3.53 (m, 2H), 3.32 (s, 3H), 2.10-2.75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

(x) Ph( 3- Br)( 5- OCHzn-(/ ?) CH( OH) C( 0)- Aze- Pab( Teoc) (x) Ph( 3- Br)( 5- OCHzn-(/ ?) CH( OH) C( 0)- Aze- Pab( Teoc)

En blanding av Ph(3-Br)(5-OCH2F)-(/?)CH(OMEM)C(0)-Aze-Pab(Teoc) (347 mg, 0,478 mmol; se trinn (ix) ovenfor) og karbontetrabromid (159 mg, 0,478 mmol) i 2-propanol (10 ml) ble tilbakeløpskokt i 1,5 timer. Blandingen ble konsentrert i vakuum deretter fordelt med H20 (20 ml) og EtOAc (3 x 30 ml). De samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:EtOH (15:1) ga sub-tittelforbindelse (59 mg, 19%) som et knusbart hvitt skum. A mixture of Ph(3-Br)(5-OCH2F)-(/?)CH(OMEM)C(0)-Aze-Pab(Teoc) (347 mg, 0.478 mmol; see step (ix) above) and carbon tetrabromide (159 mg, 0.478 mmol) in 2-propanol (10 mL) was refluxed for 1.5 h. The mixture was concentrated in vacuo then partitioned with H 2 O (20 mL) and EtOAc (3 x 30 mL). The combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl 3 :EtOH (15:1) afforded the sub-title compound (59 mg, 19%) as a crumbly white foam.

Sm.p.: 81-87°C Melting point: 81-87°C

Rf = 0,58 (9:1 CHCl3:EtOH) Rf = 0.58 (9:1 CHCl3:EtOH)

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 5 7,84 (d, / = 8 Hz, 2H), 7,40-7,48 (m, 2H) 7,18-7,30 (m, 3H), 5,80 (d, 7H-f = 53 Hz, 2H), 5,21 og 5,15 (s, 1H), 5,18-5,24 og 4,80-4,88 (m, 1H), 3,98-4,54 (m, 6H), 2,10-2,70 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s, 9H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.84 (d, / = 8 Hz, 2H), 7.40-7.48 (m, 2H) 7.18-7.30 (m, 3H), 5.80 (d, 7H-f = 53 Hz, 2H), 5.21 and 5.15 (s, 1H), 5.18-5.24 and 4.80-4.88 (m, 1H), 3.98-4.54 (m, 6H), 2.10-2.70 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s , 9H).

APCI-MS: (M + 1) = 637 m/z APCI-MS: (M + 1) = 637 m/z

(xi) Ph( 3- Br)( 5- OCH9F)-(/ ?) CH( OH) C( 0)- Aze- Pab x TFA (xi) Ph( 3- Br)( 5- OCH9F)-(/ ?) CH( OH) C( 0)- Aze- Pab x TFA

Ph(3-Br)(5-OCH2F)-(Æ)CH(OH)C(0)-Aze-Pab(Teoc) (0,073 g, 0,11 mmol; se trinn (x) ovenfor), ble oppløst i 5 ml TFA og fikk reagere i 90 min mens den ble avkjølt på et isbad. TFA ble inndampet og residuet renset ved prep RPLC med CH3CN:0,1M NH4OAC (30:70). Gjeldende fraksjoner ble inndampet og fryse tørket fra vann/acetonitril, hvilket gir 49 mg (77%) av tittelforbindelsen som dens acetatsalt. Ph(3-Br)(5-OCH2F)-(Æ)CH(OH)C(0)-Aze-Pab(Teoc) (0.073 g, 0.11 mmol; see step (x) above), was dissolved in 5 ml TFA and allowed to react for 90 min while cooling in an ice bath. The TFA was evaporated and the residue purified by prep RPLC with CH3CN:0.1M NH4OAC (30:70). Current fractions were evaporated and freeze dried from water/acetonitrile to give 49 mg (77%) of the title compound as its acetate salt.

'H-NMR (300 MHz; CD3OD) rotamerer: 8 7,8-7,7 (m, 2H), 7,54 (m, 2H), 7,37 (s, 1H, hoved rotamer), 7,33 (s, 1H, mindre rotamer), 7,25-7,1 (m, 2H), 5,75 (d, 2H), 5,22 (m, 1H, mindre rotamer), 5,18 (s, 1H, hoved rotamer), 5,11 (s, 1H, mindre rotamer), 4,80 (m, 1H, hoved rotamer), 4,6-4,4 (m, 2H), 4,37 (m, 1H, hoved rotamer), 4,16 (m, 1H, hoved rotamer), 4,1-3,9 (m, 2H, to signaler fra mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,52 (m, 1H, hoved rotamer), 2,30 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer), 1,89 (s, 3H). 1H-NMR (300 MHz; CD3OD) rotamers: δ 7.8-7.7 (m, 2H), 7.54 (m, 2H), 7.37 (s, 1H, main rotamer), 7.33 (s, 1H, minor rotamer), 7.25-7.1 (m, 2H), 5.75 (d, 2H), 5.22 (m, 1H, minor rotamer), 5.18 (s, 1H , major rotamer), 5.11 (s, 1H, minor rotamer), 4.80 (m, 1H, major rotamer), 4.6-4.4 (m, 2H), 4.37 (m, 1H, major rotamer), 4.16 (m, 1H, major rotamer), 4.1-3.9 (m, 2H, two signals from minor rotamer), 2.70 (m, 1H, minor rotamer), 2.52 (m, 1H, major rotamer), 2.30 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer), 1.89 (s, 3H).

ESI-MS+: (M+l) = 493/495 (m/z) ESI-MS+: (M+1) = 493/495 (m/z)

Eksempel 28 Example 28

Ph( 3- Br)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab x TFA Ph( 3- Br)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab x TFA

(i) 1. 3- dibrom- 5- dilfuormetoksvbenzen (i) 1. 3- dibromo- 5- difluoromethoxybenzene

Til en tarert, forseglet 350 ml rundbunnet trykk kolbe inneholdende en løsning av 3,5-dibromfenol (10,0 g, 39,7 mmol; se Eksempel 27(ii) ovenfor) i 2-propanol (100 ml) og 30% KOH (80 ml) ved -78°C ble det tilsatt klordifluormetan via bobling i 15 min gjennom septum. Septumet ble erstattet med en Teflon stopper og kolben ble deretter forseglet og fikk oppvarmes til romtemperatur hvor kolben ble veiet og bestemt å inneholde 12,0 g (138 mmol) klordifluormetan. Løsningen ble tilbakeløpskokt natten over i et oljebad satt til 80°C. Kolben ble avkjølt til romtemperatur, trykket forsiktig frigjort og innholdet fortynnet med H20 (200 ml). Det vandige laget ble ekstrahert med CHC13 (2 x 150 ml), deretter ble de samlede organiske lag tørket (NajSO^, filtrert og konsentrert i vakuum. Residuet ble renset ved Kugelrohr destillering ved 80°C ved 0,2 mm Hg, hvilker gir sub-tittelforbindelsen (9,6 g, 80%) som en klar væske. To a tared, sealed 350 mL round-bottomed pressure flask containing a solution of 3,5-dibromophenol (10.0 g, 39.7 mmol; see Example 27(ii) above) in 2-propanol (100 mL) and 30% KOH (80 mL) at -78°C was added chlorodifluoromethane via bubbling for 15 min through the septum. The septum was replaced with a Teflon stopper and the flask was then sealed and allowed to warm to room temperature where the flask was weighed and determined to contain 12.0 g (138 mmol) of chlorodifluoromethane. The solution was refluxed overnight in an oil bath set at 80°C. The flask was cooled to room temperature, the pressure carefully released and the contents diluted with H 2 O (200 mL). The aqueous layer was extracted with CHCl 3 (2 x 150 mL), then the combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. The residue was purified by Kugelrohr distillation at 80°C at 0.2 mm Hg, giving the sub-title compound (9.6 g, 80%) as a clear liquid.

'H NMR (300 MHz, CDC13) 5 7,55 (s, 1H), 7,26 (s, 2H), 6,52 (t, 7H-f = 68 Hz, 1H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.55 (s, 1H), 7.26 (s, 2H), 6.52 (t, 7H-f = 68 Hz, 1H).

(ii) l- brom- 3- difluormetoksy- 5- vinylbenzen (ii) 1-bromo-3-difluoromethoxy-5-vinylbenzene

Tri(butyl)vinyltin (10,5 g, 33,1 mmol) ble satt dråpevis til en løsning av l,3-dibrom-5-difluormetoksybenzen (9,1 g, 30,1 mmol; se trinn (i) ovenfor), Tri(butyl)vinyltin (10.5 g, 33.1 mmol) was added dropwise to a solution of 1,3-dibromo-5-difluoromethoxybenzene (9.1 g, 30.1 mmol; see step (i) above) ,

tetrakis(trifenylfosfin)palladium(0) (700 mg, 0,60 mmol) og 2,6-di-terf-butyl-4-metylfenol (spatel tip) i toluen (125 ml) under nitrogen. Blandingen ble omrørt ved 50°C natten over. Blandingen ble avkjølt til 0°C og IN NaOH (70 ml) ble tilsatt. Etter 1 time, ble blandingen ekstrahert med CH2CI2 (3 x 300 ml) deretter, ble de samlede organiske lag tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med heksaner ga sub-tittelforbindelse (5,1 g, 68%) som en fargeløs olje. tetrakis(triphenylphosphine)palladium(0) (700 mg, 0.60 mmol) and 2,6-di-tert-butyl-4-methylphenol (spatula tip) in toluene (125 mL) under nitrogen. The mixture was stirred at 50°C overnight. The mixture was cooled to 0°C and 1N NaOH (70 mL) was added. After 1 hour, the mixture was extracted with CH 2 Cl 2 (3 x 300 mL) then the combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with hexanes gave the sub-title compound (5.1 g, 68%) as a colorless oil.

<*>H NMR (300 MHz, CDCI3) 5 7,53 (s, 1H), 7,18 (s, 1H), 7,08 (s, 1H), 6,60 (dd, 7 = 6 Hz, 7 = 11 Hz, 1H), 6,57 (t, 7H-f = 68 Hz, 1H), 5,77 (d, 7 = 11 Hz, 1H), 5,36 (d, 7 = 8 Hz, 1H). <*>H NMR (300 MHz, CDCl 3 ) δ 7.53 (s, 1H), 7.18 (s, 1H), 7.08 (s, 1H), 6.60 (dd, 7 = 6 Hz, 7 = 11 Hz, 1H), 6.57 (t, 7H-f = 68 Hz, 1H), 5.77 (d, 7 = 11 Hz, 1H), 5.36 (d, 7 = 8 Hz, 1H ).

(iii) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) CH?OH (iii) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) CH?OH

2-metyl-2-propanol (150 ml), H20 (150 ml) og AD-mix-P (27,8 g) ble samlet sammen og avkjølt til 0°C. l-brom-3-difluormetoksy-5-vinylbenzen (4,6 g, 18,6 mmol; se trinn (ii) ovenfor) ble tilsatt på en gang og den heterogene oppslemningen ble kraftig omrørt ved 0°C inntil TLC viste fravær av utgangsmaterialet, deretter ble løsningen oppvarmet til romtemperatur og omrørt natten over. Reaksjonen ble stanset ved 0°C ved tilsetning av mettet natriumsulfitt (300 ml) og deretter oppvarmet til romtemperatur og omrørt i 60 min. Reaksjonsblandingen ble ekstrahert med EtOAc (3 x 200 ml). De samlede organiske ekstrakter ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (5,0 g, 95%) som en fargeløs olje som ble anvendt uten ytterligere rensning. 2-Methyl-2-propanol (150 mL), H 2 O (150 mL) and AD-mix-P (27.8 g) were combined and cooled to 0°C. 1-Bromo-3-difluoromethoxy-5-vinylbenzene (4.6 g, 18.6 mmol; see step (ii) above) was added at once and the heterogeneous slurry was vigorously stirred at 0°C until TLC showed the absence of the starting material, then the solution was warmed to room temperature and stirred overnight. The reaction was quenched at 0°C by the addition of saturated sodium sulphite (300 ml) and then warmed to room temperature and stirred for 60 min. The reaction mixture was extracted with EtOAc (3 x 200 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (5.0 g, 95%) as a colorless oil which was used without further purification.

<*>H NMR (300 MHz, CD3OD) 5 7,43 (s, 1H), 7,23 (s, 1H), 7,16 (s, 1H), 6,86 (t, 7H-f = 75 Hz, 1H), 4,64-4,67 (m, 1H), 3,54-3,59 (m, 2H). <*>H NMR (300 MHz, CD3OD) δ 7.43 (s, 1H), 7.23 (s, 1H), 7.16 (s, 1H), 6.86 (t, 7H-f = 75 Hz, 1H), 4.64-4.67 (m, 1H), 3.54-3.59 (m, 2H).

HPLC Analyse: 88,6%, 96,3% ee, ChiralPak AD Kolonne (95:5 Heks:EtOH mobil fase). HPLC Analysis: 88.6%, 96.3% ee, ChiralPak AD Column (95:5 Hex:EtOH mobile phase).

(iv) Ph( 3- Br)( 5- OCHF9)-(/ g) CH( OMEM) CH, OTBS (iv) Ph( 3- Br)( 5- OCHF9)-(/ g) CH( OMEM) CH, OTBS

Til en løsning av Ph(3-Br)(5-OCHF2)-(/?)CH(OH)CH2OH (4,9 g, 17,3 mmol; se trinn (iii) ovenfor), 4-(dimetylamino)pyridin (420 mg, 3,5 mmol) og DIPEA (11,2 g, 86,3 mmol) i vannfri CH2CI2 (250 ml) ble tilsatt dråpevis en 1,0 M løsning av tert-butyldimetylsilylklorid i CH2CI2 (20,7 ml, 20,7 mmol). Reaksjonsblandingen ble omrørt natten over ved romtemperatur. Til blandingen ble det tilsatt DIPEA (11,2 g, 86,3 mmol) og 2-metoksyetoksymetylklorid (12,9 g, 104 mmol) dråpevis. Etter 3 d, ble ytterligere 2-metoksyetoksymetylklorid (3,3 g) tilsatt og reaksjonsblandingen omrørt natten over. Blandingen ble fortynnet med vann (250 ml) og lagene ble separert. Det vandige laget ble ekstrahert med CH2CI2 (2 x 250 ml), deretter de samlede organiske lag ble tørket (Na2SC«4), filtrert og konsentrert 1 vakuum. Flash kromatografi på silikagel under eluering med Heks:EtOAc (4:1) ga sub-tittelforbindelsen (4,3 g, 51%) som en fargeløs olje. To a solution of Ph(3-Br)(5-OCHF2)-(/?)CH(OH)CH2OH (4.9 g, 17.3 mmol; see step (iii) above), 4-(dimethylamino)pyridine (420 mg, 3.5 mmol) and DIPEA (11.2 g, 86.3 mmol) in anhydrous CH 2 Cl 2 (250 mL) was added dropwise to a 1.0 M solution of tert-butyldimethylsilyl chloride in CH 2 Cl 2 (20.7 mL, 20.7 mmol). The reaction mixture was stirred overnight at room temperature. To the mixture was added DIPEA (11.2 g, 86.3 mmol) and 2-methoxyethoxymethyl chloride (12.9 g, 104 mmol) dropwise. After 3 d, additional 2-methoxyethoxymethyl chloride (3.3 g) was added and the reaction mixture was stirred overnight. The mixture was diluted with water (250 mL) and the layers were separated. The aqueous layer was extracted with CH 2 Cl 2 (2 x 250 mL), then the combined organic layers were dried (Na 2 SC 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with Hex:EtOAc (4:1) gave the sub-title compound (4.3 g, 51%) as a colorless oil.

'H NMR (300 MHz, CDCI3) 5 7,40 (s, 1H), 7,25 (s, 1H), 7,08 (s, 1H), 6,58 (t, 7H-f = 75 Hz, 1H), 4,84 (d, 7 = 7 Hz, 1H), 4,70-4,74 (m, 2H), 3,50-3,91 (m, 6H), 3,42 (s, 3H), 0,90 (s, 9H), 0,12 (s, 3H), 0,05 (s, 3H). 1 H NMR (300 MHz, CDCl 3 ) δ 7.40 (s, 1H), 7.25 (s, 1H), 7.08 (s, 1H), 6.58 (t, 7H-f = 75 Hz, 1H), 4.84 (d, 7 = 7 Hz, 1H), 4.70-4.74 (m, 2H), 3.50-3.91 (m, 6H), 3.42 (s, 3H ), 0.90 (s, 9H), 0.12 (s, 3H), 0.05 (s, 3H).

(v) Ph( 3- Br¥5- OCHF9)-(/ ?) CH( OMEM) CH90H (v) Ph( 3- Br¥5- OCHF9)-(/ ?) CH( OMEM) CH90H

Til en løsning av Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)CH2OTBS (3,3 g, 6,9 mmol; se trinn (iv) ovenfor) i THF (60 ml) ble tilsatt en 1,0 M løsning av tetrabutylammoniumfluorid i THF (9,0 ml, 9,0 mmol) ved romtemperatur. Reaksjonsblandingen ble omrørt i 45 min, blandingen ble deretter fordelt med vann (150 ml) og EtOAc (2 x 120 ml). De samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert i vakuum, hvilker gir sub-tittelforbindelsen (2,5 g, 98%) som en gul olje som ble anvendt uten ytterligere rensning. To a solution of Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)CH2OTBS (3.3 g, 6.9 mmol; see step (iv) above) in THF (60 mL) was added a 1.0 M solution of tetrabutylammonium fluoride in THF (9.0 mL, 9.0 mmol) at room temperature. The reaction mixture was stirred for 45 min, then the mixture was partitioned with water (150 mL) and EtOAc (2 x 120 mL). The combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (2.5 g, 98%) as a yellow oil which was used without further purification.

<!>H NMR (300 MHz, CD3OD) 8 7,35 (s, 1H), 7,21 (s, 1H), 7,08 (s, 1H), 6,83 (t, 7H-f = 73 Hz, 1H), 4,73 (d, 7 = 7 Hz, 1H), 4,59-4,68 (m, 2H), 3,40-380 (m, 6H), 3,26 (s, 3H). <!>H NMR (300 MHz, CD3OD) δ 7.35 (s, 1H), 7.21 (s, 1H), 7.08 (s, 1H), 6.83 (t, 7H-f = 73 Hz, 1H), 4.73 (d, 7 = 7 Hz, 1H), 4.59-4.68 (m, 2H), 3.40-380 (m, 6H), 3.26 (s, 3H ).

(vi) Ph( 3- Br)( 5- OCHF7)-(/ g) CH( OMEM) C( Q) OH (vi) Ph( 3- Br)( 5- OCHF7)-(/ g) CH( OMEM) C( Q) OH

En løsning av Ph(3-Br)(5-OCHF2)-(Æ)CH(OMEM)CH2OH (3,0 g, 8,1 mmol; se trinn (v) ovenfor) i aceton (60 ml) ble satt til en vandig 5% NaHC03 løsning (25 ml). Denne magnetisk omrørte heterogene blanding ble avkjølt til 0°C og deretter ble kaliumbromid (100 mg, 0,81 mmol) og 2,2,6,6-tetrametyl-l-piperidinyloksy, fri rest (1,3 g, 8,5 mmol) tilsatt. Natriumhypokloritt (5,25%, 19 ml) ble deretter tilsatt dråpevis over en periode på 10 min mens blandingen ble kraftig omrørt og holdt ved 0°C. Etter 1 time, ble ytterligere natriumhypokloritt (17 ml) og NaHCOs løsning (34 ml) tilsatt og omrøring ble fortsatt ved 0°C i ytterligere 4 timer. Aceton ble fjernet på en rotasjonsinndamper. Det vandige laget ble fortynnet med 10% NaHC03 løsning (30 ml) og ble vasket med Et20 (3 x 20 ml). Det vandige laget ble surgjort til pH 3,5 med 10% sitronsyre og ekstrahert med EtOAc (3 x 40 ml). De samlede EtOAc lag ble vasket med H2O (3 x 50 ml) og saltvann (50 ml) og deretter tørket (Na2S04), filtrert og konsentrert 1 vakuum, hvilker gir sub-tittelforbindelsen (2,1 g, 66%) som en fargeløs olje som ble anvendt uten ytterligere rensning. A solution of Ph(3-Br)(5-OCHF2)-(Æ)CH(OMEM)CH2OH (3.0 g, 8.1 mmol; see step (v) above) in acetone (60 mL) was added an aqueous 5% NaHCO 3 solution (25 mL). This magnetically stirred heterogeneous mixture was cooled to 0°C and then potassium bromide (100 mg, 0.81 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy, free residue (1.3 g, 8.5 mmol) added. Sodium hypochlorite (5.25%, 19 mL) was then added dropwise over a period of 10 min while the mixture was vigorously stirred and maintained at 0°C. After 1 hour, additional sodium hypochlorite (17 mL) and NaHCO 3 solution (34 mL) were added and stirring was continued at 0°C for an additional 4 hours. Acetone was removed on a rotary evaporator. The aqueous layer was diluted with 10% NaHCO 3 solution (30 mL) and was washed with Et 2 O (3 x 20 mL). The aqueous layer was acidified to pH 3.5 with 10% citric acid and extracted with EtOAc (3 x 40 mL). The combined EtOAc layers were washed with H 2 O (3 x 50 mL) and brine (50 mL) then dried (Na 2 SO 4 ), filtered and concentrated in vacuo to give the sub-title compound (2.1 g, 66%) as a colorless oil that was used without further purification.

'H NMR (300 MHz, CD3OD) 8 7,51 (s, 1H), 7,32 (s, 1H), 7,24 (s, 1H), 6,88 (t, 7H-f = 73 Hz, 1H), 5,21 (s, 1H), 4,84 (d, 7 = 7 Hz, 1H), 4,76 (d, 7 = 7 Hz, 1H), 3,62-3,80 (m, 2H), 3,48-3,52 (m, 2H), 3,32 (s, 3H). 1 H NMR (300 MHz, CD 3 OD) δ 7.51 (s, 1H), 7.32 (s, 1H), 7.24 (s, 1H), 6.88 (t, 7H-f = 73 Hz, 1H), 5.21 (s, 1H), 4.84 (d, 7 = 7 Hz, 1H), 4.76 (d, 7 = 7 Hz, 1H), 3.62-3.80 (m, 2H), 3.48-3.52 (m, 2H), 3.32 (s, 3H).

(vii) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OMEM) C( 0)- Aze- Pab( Teoc) (vii) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OMEM) C( 0)- Aze- Pab( Teoc)

Til en løsning av Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (1,0 g, 2,62 mmol; se trinn (vi) ovenfor) i DMF (50 ml) under nitrogen ved 0°C ble det tilsatt HAze-Pab(Teoc)»HCl (1,5 g, 3,38 mmol), PyBOP (1,5 g, 2,9 mmol) og DIPEA (840 mg, 6,50 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet kromatografert på silikagel under eluering med CHCl3:EtOH (15:1), hvilker gir sub-tittelforbindelsen (1,1 g, 59%) som et knusbart hvitt skum. To a solution of Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (1.0 g, 2.62 mmol; see step (vi) above) in DMF ( 50 ml) under nitrogen at 0°C was added HAze-Pab(Teoc)»HCl (1.5 g, 3.38 mmol), PyBOP (1.5 g, 2.9 mmol) and DIPEA (840 mg, 6.50 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed on silica gel eluting with CHCl 3 :EtOH (15:1) to give the sub-title compound (1.1 g, 59%) as a breakable white foam.

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,79-7,83 (m, 2H), 7,26-7,52 (m, 5H), 6,94 og 6,91 (t, 7H-f = 73 Hz, 1H), 5,27 og 5,07 (s, 1H), 5,20-5,23 og 4,80-4,88 (m, 1H), 4,01-4,79 (m, 8H), 3,60-3,71 (m, 2H), 3,40-3,53 (m, 2H), 3,32 (s, 3H), 2,10-2,75 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s, 9H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.79-7.83 (m, 2H), 7.26-7.52 (m, 5H), 6.94 and 6.91 (t, 7H-f = 73 Hz, 1H), 5.27 and 5.07 (s, 1H), 5.20-5.23 and 4.80-4.88 (m, 1H), 4.01 -4.79 (m, 8H), 3.60-3.71 (m, 2H), 3.40-3.53 (m, 2H), 3.32 (s, 3H), 2.10-2 .75 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s, 9H).

(viii) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( Teoc) (viii) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( Teoc)

En blanding av Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(Teoc) (369 mg, 0,496 mmol; se trinn (vii) ovenfor) og karbontetrabromid (165 mg, 0,496 mmol) i 2-propanol (10 ml) ble tilbakeløpskokt i 12 timer. Blandingen ble konsentrert 1 vakuum, deretter fordelt med H20 (15 ml) og EtOAc (5 x 20 ml). De samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:EtOH (15:1) ga sub-tittelforbindelsen (134 mg, 41%) som et knusbart hvitt skum. A mixture of Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(Teoc) (369 mg, 0.496 mmol; see step (vii) above) and carbon tetrabromide (165 mg, 0.496 mmol) in 2-propanol (10 mL) was refluxed for 12 h. The mixture was concentrated in vacuo, then partitioned with H 2 O (15 mL) and EtOAc (5 x 20 mL). The combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl 3 :EtOH (15:1) afforded the sub-title compound (134 mg, 41%) as a crushable white foam.

Sm.p.: 92-98°C Melting point: 92-98°C

Rf = 0,37 (9:1 CHCl3:EtOH) Rf = 0.37 (9:1 CHCl3:EtOH)

lU NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,80-7,86 (m, 2H), 7,40-7,48 (m, 2H) 7,10-7,33 (m, 3H), 6,92 og 6,88 (t, 7H-f = 73 Hz, 1H), 5,18 og 5,11 (s, 1H), 5,18-5,24 og 4,76-4,80 (m, 1H), 3,98-4,54 (m, 6H), 2,10-2,70 (m, 2H), 1,05-1,11 (m, 2H), 0,08 (s, 9H). 1U NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.80-7.86 (m, 2H), 7.40-7.48 (m, 2H) 7.10-7.33 (m, 3H ), 6.92 and 6.88 (t, 7H-f = 73 Hz, 1H), 5.18 and 5.11 (s, 1H), 5.18-5.24 and 4.76-4.80 (m, 1H), 3.98-4.54 (m, 6H), 2.10-2.70 (m, 2H), 1.05-1.11 (m, 2H), 0.08 (s , 9H).

APCI-MS: (M + 1) = 655 m/z APCI-MS: (M + 1) = 655 m/z

(ix) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA (ix) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab x TFA

Ph(3-Br)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(Teoc) (0,081 g, 0,124 mmol; se trinn (viii) ovenfor), ble oppløst i 5 ml TFA og fikk reagere i 80 min mens den ble avkjølt på et isbad. TFA ble inndampet og residuet renset ved prep RPLC med CH3CN:0,1M NH4OAC (30:70). Gjeldende fraksjoner ble inndampet og fryse tørket fra vann/acetonitril, hvilket gir 59 mg (83%) av tittelforbindelsen som dens acetatsalt. Ph(3-Br)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(Teoc) (0.081 g, 0.124 mmol; see step (viii) above), was dissolved in 5 mL TFA and allowed to react for 80 min while it was cooled in an ice bath. The TFA was evaporated and the residue purified by prep RPLC with CH3CN:0.1M NH4OAC (30:70). Current fractions were evaporated and freeze dried from water/acetonitrile to give 59 mg (83%) of the title compound as its acetate salt.

'H-NMR (300 MHz; CD3OD) rotamerer: 8 7,8-7,7 (m, 2H), 7,6-7,4 (m, 3H), 7,3-7,2 (m, 2H), 6,89 (t, 1H, hoved rotamer), 6,87 (t, 1H, mindre rotamer), 5,23 (m, 1H, mindre rotamer), 5,21 (s, 1H, hoved rotamer), 5,13 (s, 1H, mindre rotamer), 4,80 (m, 1H, hoved rotamer), 4,6-4,4 (m, 2H), 4,38 (m, 1H, hoved rotamer), 4,20 (m, 1H, hoved rotamer), 4,1-3,9 (m, 2H, to signaler fra mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,54 (m, 1H, hoved rotamer), 2,29 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer), 1,89 (s, 3H). <13>C-NMR (75 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) 8 172,0,171,7, 167,0. 1H-NMR (300 MHz; CD 3 OD) rotamers: δ 7.8-7.7 (m, 2H), 7.6-7.4 (m, 3H), 7.3-7.2 (m, 2H ), 6.89 (t, 1H, major rotamer), 6.87 (t, 1H, minor rotamer), 5.23 (m, 1H, minor rotamer), 5.21 (s, 1H, major rotamer), 5.13 (s, 1H, minor rotamer), 4.80 (m, 1H, major rotamer), 4.6-4.4 (m, 2H), 4.38 (m, 1H, major rotamer), 4 .20 (m, 1H, major rotamer), 4.1-3.9 (m, 2H, two signals from minor rotamer), 2.70 (m, 1H, minor rotamer), 2.54 (m, 1H, major rotamer), 2.29 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer), 1.89 (s, 3H). <13>C-NMR (75 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms) δ 172.0, 171.7, 167.0.

MS (m/z) 511/513 (M+l)<+>MS (m/z) 511/513 (M+1)<+>

Eksempel 29 Example 29

Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( OMe) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( OMe)

(i) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OMEM) C( 0)- Aze- Pab( OMe) (i) Ph( 3- Br)( 5- OCHF2)-(/ g) CH( OMEM) C( 0)- Aze- Pab( OMe)

Til en løsning av Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (957 mg, 2,48 mmol; se Eksempel 28(vi) ovenfor) i DMF (30 ml) under nitrogen ved 0°C ble det tilsatt HAze-Pab(OMe)-2HCl (1,1 g, 3,2 mmol), PyBOP (1,4 g, 2,7 mmol) og DIPEA (804 mg, 6,2 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 2 timer og deretter ved romtemperatur natten over. Blandingen ble konsentrert i vakuum og residuet kromatografert to ganger på silikagel, under eluering først med CHCl3:EtOH (9:1) og deretter med EtOAc:EtOH (15:1), hvilker gir sub-tittelforbindelsen (1,1 g, 72%) som et knusbart hvitt skum. To a solution of Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)OH (957 mg, 2.48 mmol; see Example 28(vi) above) in DMF (30 ml) under nitrogen at 0°C was added HAze-Pab(OMe)-2HCl (1.1 g, 3.2 mmol), PyBOP (1.4 g, 2.7 mmol) and DIPEA (804 mg, 6 .2 mmol). The reaction mixture was stirred at 0°C for 2 hours and then at room temperature overnight. The mixture was concentrated in vacuo and the residue chromatographed twice on silica gel, eluting first with CHCl3:EtOH (9:1) and then with EtOAc:EtOH (15:1) to give the sub-title compound (1.1 g, 72% ) as a crushable white foam.

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,59-7,65 (m, 2H), 7,20-7,55 (m, 5H), 6,95 og 6,91 (t, 7H-f = 73 Hz, 1H), 5,27 og 5,07 (s, 1H), 5,18-5,23 og 4,75-4,84 (m, 1H), 3,87-4,89 (m, 6H), 3,84 (s, 3H), 3,60-3,71 (m, 2H), 3,40-3,53 (m, 2H), 3,32 (s, 3H), 2,10-2,75 (m, 2H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.59-7.65 (m, 2H), 7.20-7.55 (m, 5H), 6.95 and 6.91 (t, 7H-f = 73 Hz, 1H), 5.27 and 5.07 (s, 1H), 5.18-5.23 and 4.75-4.84 (m, 1H), 3.87 -4.89 (m, 6H), 3.84 (s, 3H), 3.60-3.71 (m, 2H), 3.40-3.53 (m, 2H), 3.32 (s , 3H), 2.10-2.75 (m, 2H).

(ii) Ph( 3- Br)( 5- OCHF7)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe) (ii) Ph( 3- Br)( 5- OCHF7)-(/ ?) CH( OH) C( 0)- Aze- Pab( OMe)

En blanding av Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(OMe) (1,1 g, 1,8 mmol; se trinn (i) ovenfor) og karbontetrabromid (583 mg, 1,8 mmol) i 2-propanol (30 ml) ble tilbakeløpskokt i 2,5 d. I løpet av denne tiden ble ytterligere karbontetrabromid (5 porsjoner av 50 mg i intervaller for ytterligere 0,90 mmol) satt til for å sikre fullføring av reaksjonen. Blandingen ble konsentrert i vakuum, deretter fordelt med H20 (50 ml) og EtOAc (5 x 25 ml). De samlede organiske lag ble tørket (Na2S04), filtrert og konsentrert i vakuum. Flash kromatografi på silikagel under eluering med CHCl3:EtOH (15:1) ga tittelforbindelsen (460 mg, 50%) som et knusbart hvitt skum. A mixture of Ph(3-Br)(5-OCHF2)-(/?)CH(OMEM)C(0)-Aze-Pab(OMe) (1.1 g, 1.8 mmol; see step (i) above) and carbon tetrabromide (583 mg, 1.8 mmol) in 2-propanol (30 mL) were refluxed for 2.5 d. During this time, additional carbon tetrabromide (5 portions of 50 mg at intervals for an additional 0.90 mmol) was added to ensure completion of the reaction. The mixture was concentrated in vacuo, then partitioned with H 2 O (50 mL) and EtOAc (5 x 25 mL). The combined organic layers were dried (Na 2 SO 4 ), filtered and concentrated in vacuo. Flash chromatography on silica gel eluting with CHCl 3 :EtOH (15:1) afforded the title compound (460 mg, 50%) as a crushable white foam.

Sm.p.: 71-75°C Melting point: 71-75°C

Rf = 0,63 (9:1 CHCl3:EtOH) Rf = 0.63 (9:1 CHCl3:EtOH)

<*>H NMR (300 MHz, CD3OD, kompleks blanding av rotamerer) 8 7,59 (d, 7 = 8 Hz, 2H), 7,20-7,54 (m, 5H), 6,90 og 6,87 (t, 7H-f = 73 Hz, 1H), 5,18 og 5,11 (s, 1H), 4,76-4,80 (m, 1H), 3,98-4,54 (m, 4H), 3,82 (s, 3H), 2,10-2,70 (m, 2H). <*>H NMR (300 MHz, CD3OD, complex mixture of rotamers) δ 7.59 (d, 7 = 8 Hz, 2H), 7.20-7.54 (m, 5H), 6.90 and 6, 87 (t, 7H-f = 73 Hz, 1H), 5.18 and 5.11 (s, 1H), 4.76-4.80 (m, 1H), 3.98-4.54 (m, 4H), 3.82 (s, 3H), 2.10-2.70 (m, 2H).

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,5,172,1,171,6,154,1. <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 172.5,172,1,171,6,154,1.

APCI-MS: (M + 1) = 542 m/z APCI-MS: (M + 1) = 542 m/z

Eksempel 30 Example 30

Ph( 3- Cl)( 5- OCH7CHF7)- r/ ?) CH( OH) C( 0)- Aze- Pab( OH) Ph( 3- Cl)( 5- OCH7CHF7)- r/ ?) CH( OH) C( 0)- Aze- Pab( OH)

(i) Ph( 3- Cl)( 5- OCH9CHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( Z) (i) Ph( 3- Cl)( 5- OCH9CHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( Z)

Boc-Aze-Pab(Z) (se internasjonal patentsøknad WO 97/02284, 92 mg, 0,197 mmol) ble oppløst i 10 ml EtOAc mettet med HCl(g) og fikk reagere i 10 min. Løsningsmidlet ble avdampet og residuet ble blandet med Ph(3-Cl)(5-OCH2CHF2)-r/?;CH(OH)C(0)OH (50 mg, 0,188 mmol; se Eksempel 17(v) ovenfor), PyBOP (109 mg, 0,209 mmol) og til slutt diisopropyletylamin (96 mg, 0,75 mmol) i 2 ml DMF. Blandingen ble omrørt i 2 timer og deretter hellet i 50 ml vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fasen ble vasket med vann, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel med EtOAc:MeOH (9:1). Utbytte: 100 mg (87%). Boc-Aze-Pab(Z) (see International Patent Application WO 97/02284, 92 mg, 0.197 mmol) was dissolved in 10 ml of EtOAc saturated with HCl(g) and allowed to react for 10 min. The solvent was evaporated and the residue was mixed with Ph(3-Cl)(5-OCH2CHF2)-r/?;CH(OH)C(0)OH (50 mg, 0.188 mmol; see Example 17(v) above), PyBOP (109 mg, 0.209 mmol) and finally diisopropylethylamine (96 mg, 0.75 mmol) in 2 mL DMF. The mixture was stirred for 2 h and then poured into 50 mL of water and extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc:MeOH (9:1). Yield: 100 mg (87%).

<!>H NMR (300 MHz, CD3OD blanding av rotamerer) 8 7,85-7,75 (m, 2H), 7,45-7,25 (m, 7H), 7,11 (m, 1H, hoved rotamer), 7,08 (m, 1H, mindre rotamer), 7,05-6,9 (m, 2H), 6,13 (bt, 1H), 5,25-5,05 (m, 3H), 4,77 (m, 1H, delvis hidden by CD3OH signal), 4,5-3,9 (m, 7H), 2,64 (m, 1H, mindre rotamer), 2,47 (m, 1H, hoved rotamer), 2,25 (m, 1H, hoved rotamer), 2,13 (m, 1H, mindre rotamer) <!>H NMR (300 MHz, CD3OD mixture of rotamers) δ 7.85-7.75 (m, 2H), 7.45-7.25 (m, 7H), 7.11 (m, 1H, main rotamer), 7.08 (m, 1H, minor rotamer), 7.05-6.9 (m, 2H), 6.13 (bt, 1H), 5.25-5.05 (m, 3H), 4.77 (m, 1H, partially hidden by CD3OH signal), 4.5-3.9 (m, 7H), 2.64 (m, 1H, minor rotamer), 2.47 (m, 1H, major rotamer ), 2.25 (m, 1H, major rotamer), 2.13 (m, 1H, minor rotamer)

(ii) Ph( 3- Cl)( 5- OCH9CHF7)- r/ g) CH( OH) C( Q)- Aze- Pab( OH) (ii) Ph( 3- Cl)( 5- OCH9CHF7)- r/ g) CH( OH) C( Q)- Aze- Pab( OH)

Hydroksylamin-hydroklorid (65 mg, 0,94 mmol) og trietylamin (0,319 g, 3,16 mmol) ble blandet i 8 ml THF og ultralydbehandlet i 1 time ved 40°C. Ph(3-Cl)(5-OCH2CHF2)-(Æ;CH(OH)C(0)-Aze-Pab(Z) (96 mg, 0,156 mmol; se trinn (i) ovenfor) ble tilsatt med 8 ml mer av THF. Blandingen ble omrørt ved 40°C i 4,5 dager. Løsningsmidlet ble avdampet og råproduktet ble renset ved preparativ RPLC med CH3CN:0,1M NH4OAC (40:60). Utbytte: 30 mg (38%). Renhet: 99%. Hydroxylamine hydrochloride (65 mg, 0.94 mmol) and triethylamine (0.319 g, 3.16 mmol) were mixed in 8 mL of THF and sonicated for 1 hour at 40°C. Ph(3-Cl)(5-OCH2CHF2)-(Æ;CH(OH)C(0)-Aze-Pab(Z) (96 mg, 0.156 mmol; see step (i) above) was added with 8 mL more of THF. The mixture was stirred at 40°C for 4.5 days. The solvent was evaporated and the crude product was purified by preparative RPLC with CH3CN:0.1M NH4OAC (40:60). Yield: 30 mg (38%). Purity: 99%.

<]>H NMR (300 MHz, CD3OD blanding av rotamerer) 8 7,6-7,55 (m, 2H), 7,35-7,3 (m, 2H), 7,12 (m, 1H, hoved rotamer), 7,09 (m, 1H, mindre rotamer), 7,05-6,9 (m, 2H), 6,15 (triplett av multippeller, 1H), 5,15 (m, 1H, mindre rotamer), 5,13 (s, 1H, hoved rotamer), 5,08 (s, 1H, mindre rotamer), 4,77 (m, 1H, hoved rotamer), 4,5-4,2 (m, 5H), 4,08 (m, 1H, hoved rotamer), 3,97 (m, 1H, mindre rotamer), 2,66 (m, 1H, mindre rotamer), 2,50 (m, 1H hoved rotamer), 2,27 (m, 1H, hoved rotamer), 2,14 (m, 1H, mindre rotamer). <]>H NMR (300 MHz, CD3OD mixture of rotamers) δ 7.6-7.55 (m, 2H), 7.35-7.3 (m, 2H), 7.12 (m, 1H, main rotamer), 7.09 (m, 1H, minor rotamer), 7.05-6.9 (m, 2H), 6.15 (triplet of multiples, 1H), 5.15 (m, 1H, minor rotamer) , 5.13 (s, 1H, major rotamer), 5.08 (s, 1H, minor rotamer), 4.77 (m, 1H, major rotamer), 4.5-4.2 (m, 5H), 4.08 (m, 1H, major rotamer), 3.97 (m, 1H, minor rotamer), 2.66 (m, 1H, minor rotamer), 2.50 (m, 1H major rotamer), 2.27 (m, 1H, major rotamer), 2.14 (m, 1H, minor rotamer).

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 172,8,172,2,171,4,159,1,158,9,154,2. <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 172,8,172,2,171,4,159,1,158,9,154,2.

APCI-MS: (M + 1) = 497/499 m/z APCI-MS: (M + 1) = 497/499 m/z

Eksempel 31 Example 31

Ph( 3- Cl)( 5- OCH7CH7F)-(/ g) CH( OH) C( Q)- Aze- Pab( OH) Ph( 3- Cl)( 5- OCH7CH7F)-(/ g) CH( OH) C( Q)- Aze- Pab( OH)

(i) Ph( 3- Cl)( 5- OCH9CH7FW/ ?) CH( OH) C( 0)- Aze- Pab( Z) (i) Ph( 3- Cl)( 5- OCH9CH7FW/ ?) CH( OH) C( 0)- Aze- Pab( Z)

Boc-Aze-Pab(Z) (130 mg, 0,279 mmol) ble oppløst i 15 ml EtOAc mettet med HCl(g) og fikk reagere i 10 min. Løsningsmidlet ble avdampet og residuet ble blandet med Ph(3-Cl)(5-OCH2CH2F)-r/?>)CH(OH)-C(0)OH (63 mg, 0,188 mmol; se Eksempel 21(v) ovenfor) i 3 ml DMF, PyBOP (147 mg, 0,279 mmol) og til slutt diisopropyletylamin (134 mg, 1,03 mmol). Blandingen ble omrørt i 130 min og deretter hellet i 75 ml vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel med EtOAc/MeOH = 95/5. Utbytte: 119 mg (79%). Boc-Aze-Pab(Z) (130 mg, 0.279 mmol) was dissolved in 15 mL of EtOAc saturated with HCl(g) and allowed to react for 10 min. The solvent was evaporated and the residue was mixed with Ph(3-Cl)(5-OCH2CH2F)-r/?>)CH(OH)-C(0)OH (63 mg, 0.188 mmol; see Example 21(v) above) in 3 mL DMF, PyBOP (147 mg, 0.279 mmol) and finally diisopropylethylamine (134 mg, 1.03 mmol). The mixture was stirred for 130 min and then poured into 75 mL of water and extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc/MeOH = 95/5. Yield: 119 mg (79%).

<*>H NMR (400 MHz, CDC13) 88,06 (bt, 1H), 7,67 (d, 2H), 7,45-7,25 (m, 5H), 7,18 (d, 2H), 6,89 (m, 1H), 6,84 (m, 1H), 6,76 (m, 1H), 5,16 (s, 2H), 4,84 (s, 1H), 4,79 (m, 1H), 4,66 (dublett av multippeller, 2H), 4,4-4,3 (m, 2H), 4,10 (dublett av multippeller, 2H), 4,02 (m, 1H), 3,67 (m, 1H), 2,46 (m, 1H), 2,28 (m, 1H). <*>H NMR (400 MHz, CDCl 3 ) 88.06 (bt, 1H), 7.67 (d, 2H), 7.45-7.25 (m, 5H), 7.18 (d, 2H) , 6.89 (m, 1H), 6.84 (m, 1H), 6.76 (m, 1H), 5.16 (s, 2H), 4.84 (s, 1H), 4.79 ( m, 1H), 4.66 (doublet of multiples, 2H), 4.4-4.3 (m, 2H), 4.10 (doublet of multiples, 2H), 4.02 (m, 1H), 3 .67 (m, 1H), 2.46 (m, 1H), 2.28 (m, 1H).

fii) Ph( 3- Cl)( 5- OCH, CH7FW/ g) CH( OH) C( Q)- Aze- Pab( OH) fii) Ph( 3- Cl)( 5- OCH, CH7FW/ g) CH( OH) C( Q)- Aze- Pab( OH)

Hydroksylamin-hydroklorid (80 mg, 1,16 mmol) og trietylamin (0,392 g, 3,87 mmol) ble blandet i 9 ml THF og ultralydbehandlet i 1 time ved 40°C. Ph(3-Cl)(5-OCH2CH2F)-(7?jCH(OH)C(0)-Aze-Pab(Z) (96 mg, 0,156 mmol; se trinn (i) ovenfor) ble tilsatt med 9 ml mer av THF. Blandingen ble omrørt ved 40°C i 48 timer og 3 dager ved romtemperatur. Løsningsmidlet ble avdampet og råproduktet ble renset ved preparativ RPLC med CH3CN:0,1M NH4OAC (30:70). Utbytte: 72 mg (78%). Renhet: 100%. Hydroxylamine hydrochloride (80 mg, 1.16 mmol) and triethylamine (0.392 g, 3.87 mmol) were mixed in 9 mL of THF and sonicated for 1 hour at 40°C. Ph(3-Cl)(5-OCH2CH2F)-(7?jCH(OH)C(0)-Aze-Pab(Z) (96 mg, 0.156 mmol; see step (i) above) was added with 9 mL more of THF. The mixture was stirred at 40°C for 48 hours and 3 days at room temperature. The solvent was evaporated and the crude product was purified by preparative RPLC with CH3CN:0.1M NH4OAC (30:70). Yield: 72 mg (78%) .Purity: 100%.

'H NMR (400 MHz, CD3OD, blanding av rotamerer) 87,6-7,55 (m, 2H), 7,35-7,25 (m, 4H), 7,07 (m, 1H, hoved rotamer), 7,04 (m, 1H, mindre rotamer), 7,0-6,9 (M, 2 timer), 5,12 (m, 1H, mindre rotamer), 5,08 (s, 1H, mindre rotamer), 5,04 (s, 1H), 4,78 (m, 1H, hoved rotamer), 4,68 (dublett av multippelts, 2 H), 4,5-4,25 (m, 3H), 4,20 (dublett av multippeller, 2H) 4,06 (m, 1H, hoved rotamer), 3,97 (m, 1H, mindre rotamer), 2,65 (m, 1H, mindre rotamer), 2,48 (m, 1H hoved rotamer), 2,27 (m, 1H, hoved rotamer), 2,14 (m, 1H, mindre rotamer) 1 H NMR (400 MHz, CD 3 OD, mixture of rotamers) 87.6-7.55 (m, 2H), 7.35-7.25 (m, 4H), 7.07 (m, 1H, major rotamer) , 7.04 (m, 1H, minor rotamer), 7.0-6.9 (M, 2 h), 5.12 (m, 1H, minor rotamer), 5.08 (s, 1H, minor rotamer) , 5.04 (s, 1H), 4.78 (m, 1H, main rotamer), 4.68 (doublet of multiplets, 2H), 4.5-4.25 (m, 3H), 4.20 (doublet of multiples, 2H) 4.06 (m, 1H, major rotamer), 3.97 (m, 1H, minor rotamer), 2.65 (m, 1H, minor rotamer), 2.48 (m, 1H major rotamer), 2.27 (m, 1H, major rotamer), 2.14 (m, 1H, minor rotamer)

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 5 172,3,171,5,159,8,154,3 <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 5 172,3,171,5,159,8,154,3

APCI-MS: (M + 1) = 479/481 m/z APCI-MS: (M + 1) = 479/481 m/z

Eksempel 32 Example 32

Ph( 3- Cl)( 5- OCHF, W/ g) CH( OHVC( 0)- Pro- Pab Ph( 3- Cl)( 5- OCHF, W/ g) CH( OHVC( 0)- Pro- Pab

(i) Boc- Pro- Pab( Teoc) (i) Boc-Pro-Pab(Teoc)

Boc-Pro-Pab(Z) (se internasjonal patentsøknad WO 97/02284,15,0 g, 0,0321 mol) ble oppløst i 150 ml etanol og 200 mg 10% Pd/C (50% fuktighet) ble tilsatt. Blandingen ble omrørt og hydrogenen ved atmosfærisk trykk i 2 timer, filtrert gjennom Hyflo og konsentrert. Produktet ble anvendt uten ytterligere rensning. Av dette produktet ble det tatt 10 g (0,029 mol), som ble oppløst i 300 ml THF. Teoc-p-nitrofenylkarbonat (10 g, 0,035 mol) ble tilsatt. En løsning av kaliumkarbonat (5,2 g, 0,038 mol) i 50 ml vann ble tilsatt over 3 min og den resulterende løsning ble omrørt i 3 dager, konsentrert og resten ble ekstrahert med EtOAc tre ganger. De samlede organiske lag ble vasket med vann, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel ved anvendelse av metylenklorid:aceton (4:1). Utbytte: 9,8 g (69%). Boc-Pro-Pab(Z) (see International Patent Application WO 97/02284, 15.0 g, 0.0321 mol) was dissolved in 150 ml of ethanol and 200 mg of 10% Pd/C (50% humidity) was added. The mixture was stirred and the hydrogen at atmospheric pressure for 2 hours, filtered through Hyflo and concentrated. The product was used without further purification. Of this product, 10 g (0.029 mol) were taken, which was dissolved in 300 ml of THF. Teoc-p-nitrophenyl carbonate (10 g, 0.035 mol) was added. A solution of potassium carbonate (5.2 g, 0.038 mol) in 50 mL of water was added over 3 min and the resulting solution was stirred for 3 days, concentrated and the residue was extracted with EtOAc three times. The combined organic layers were washed with water, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel using methylene chloride:acetone (4:1). Yield: 9.8 g (69%).

(ii) Ph( 3- Cl)( 5- OCHF9)- r/ gJCH( OH)- C( Q)- Pro- Pab( Teoc) (ii) Ph( 3- Cl)( 5- OCHF9)- r/ gJCH( OH)- C( Q)- Pro- Pab( Teoc)

Boc-Pro-Pab(Teoe) (107 mg, 0,218 mmol; se trinn (i) ovenfor) ble oppløst i 10 ml EtOAc mettet med HCl(g) og fikk reagere i 10 min. Løsningsmidlet ble avdampet og residuet ble blandet med Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)OH (50 mg, 0,198 mmol; se Eksempel l(viii) ovenfor) i 3 ml DMF, PyBOP (115 mg, 0,218 mmol) og til slutt diisopropyletylamin (104 mg, 0,80 mmol). Blandingen ble omrørt i 2 timer og deretter hellet i 75 ml vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel med EtOAc:MeOH (95:5). Utbytte: 89 mg (72%). Boc-Pro-Pab(Teoe) (107 mg, 0.218 mmol; see step (i) above) was dissolved in 10 mL EtOAc saturated with HCl(g) and allowed to react for 10 min. The solvent was evaporated and the residue was mixed with Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)OH (50 mg, 0.198 mmol; see Example 1(viii) above) in 3 mL DMF, PyBOP (115 mg, 0.218 mmol) and finally diisopropylethylamine (104 mg, 0.80 mmol). The mixture was stirred for 2 h and then poured into 75 mL of water and extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc:MeOH (95:5). Yield: 89 mg (72%).

<*>H NMR (400 MHz, CDC13) 87,54 (bt, 1H), 7,47 (d, 2H), 7,12 (m, 1H), 7,08 (d, 2H), 7,02 (m, 1H), 6,95 (m, 1H), 6,50 (t, 1H), 5,21 (s, 1H), 4,42 (m, 1H), 4,35-4,15 (m, 3H), 3,59 (m, 1H), 2,94 (m, 1H), 2,1-1,7 (m, 4H), 1,06 (m, 2H), 0,04 (s, 9H). <*>H NMR (400 MHz, CDCl 3 ) 87.54 (bt, 1H), 7.47 (d, 2H), 7.12 (m, 1H), 7.08 (d, 2H), 7.02 (m, 1H), 6.95 (m, 1H), 6.50 (t, 1H), 5.21 (s, 1H), 4.42 (m, 1H), 4.35-4.15 ( m, 3H), 3.59 (m, 1H), 2.94 (m, 1H), 2.1-1.7 (m, 4H), 1.06 (m, 2H), 0.04 (s , 9H).

(iii) Ph( 3- Cl)( 5- OCHF2)- r/ g) CH( OH)- C( 0)- Pro- Pab x TFA (iii) Ph( 3- Cl)( 5- OCHF2)- r/ g) CH( OH)- C( 0)- Pro- Pab x TFA

Ph(3-Cl)(5-OCHF2)-WCH(OH)C(0)-Pro-Pab(Teoc) (85 mg, 0,136 mmol; se trinn (ii) ovenfor) ble oppløst i 1 ml metylenklorid og avkjølt på et isbad. TFA (4 ml) ble tilsatt og reaksjonsblandingen ble omrørt i 90 min. TFA ble inndampet og residuet ble frysetørket fra vann og acetonitril. Utbytte: 79 mg (92%). Renhet: 94%. Ph(3-Cl)(5-OCHF2)-WCH(OH)C(0)-Pro-Pab(Teoc) (85 mg, 0.136 mmol; see step (ii) above) was dissolved in 1 mL of methylene chloride and cooled on an ice bath. TFA (4 mL) was added and the reaction mixture was stirred for 90 min. The TFA was evaporated and the residue was freeze-dried from water and acetonitrile. Yield: 79 mg (92%). Purity: 94%.

<!>H NMR (400 MHz, CD3OD blanding av rotamerer) 87,85-7,7 (m, 2H), 7,58 (d, 2H, hoved rotamer), 7,47 (d, 2H, mindre rotamer), 7,35 (m, 1H, hoved rotamer), 7,27 (m, 1H, mindre rotamer), 7,2,7,1 (m, 2H), 6,88 (t, 1H), 5,38 (s, 1H, hoved rotamer), 5,22 (s, 1H, mindre rotamer), 4,58 (d, 1H), 4,5-4,2 (m, 2H), 3,8-3,5 (m, 1H), 3,35 (m, 1H), 2,2-1,8 (m, 4H). <13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer) 8 173,6,171,1, 167,0. <!>H NMR (400 MHz, CD3OD mixture of rotamers) 87.85-7.7 (m, 2H), 7.58 (d, 2H, major rotamer), 7.47 (d, 2H, minor rotamer) , 7.35 (m, 1H, major rotamer), 7.27 (m, 1H, minor rotamer), 7.2,7.1 (m, 2H), 6.88 (t, 1H), 5.38 (s, 1H, major rotamer), 5.22 (s, 1H, minor rotamer), 4.58 (d, 1H), 4.5-4.2 (m, 2H), 3.8-3.5 (m, 1H), 3.35 (m, 1H), 2.2-1.8 (m, 4H). <13>C-NMR (100 MHz; CD 3 OD): (carbonyl and/or amidine carbon atoms) δ 173.6, 171.1, 167.0.

APCI-MS: (M + 1) = 481/483 m/z APCI-MS: (M + 1) = 481/483 m/z

Eksempel 33 Example 33

Ph( 3- Cl)( 5- OCHF2)-( 7g) CH( OH)- C( Q)- Pro- Pab( OMe) Ph( 3- Cl)( 5- OCHF2)-( 7g) CH( OH)- C( Q)- Pro- Pab( OMe)

(i) 4- Azidometvl- N- metoksv- benzamidin (i) 4-Azidomethyl-N-methoxybenzamidine

4-Azidometylbenzonitril (17,3 g, 0,109 mol; Nishiyama et al; Chem. Lett. (1982) 1477) ble oppløst i 500 ml toluen og 200 ml absolutt etanol. Løsningen ble avkjølt til -10°C og HCl(g) ble boblet gjennom inntil metning. Blandingen ble holdt i kjøleskap i 2 dager når mesteparten av løsningsmidlene ble avdampet. Dietyleter ble tilsatt og ble dekantert fra. Produktet ble gjenoppløst i en løsning av O-metylhydroksylamin (10,5 g, 0,125 mol) og trietylamin (56 ml) i 200 ml metanol. Blandingen fikk stå i 3 dager mens metanolen ble inndampet med tilsetning av EtOAc. Den organiske fasen ble vasket med vann, fortynnet HOAc og vandig natriumbikarbonat, tørket (Na2S04) og fortynnet med mer EtOAc til et totalt volum på 500 ml. En prøve av 25 ml ble inndampet til tørrhet. Resten ble 932 mg. Total utbytte: 18,6 g (83%). 4-Azidomethylbenzonitrile (17.3 g, 0.109 mol; Nishiyama et al; Chem. Lett. (1982) 1477) was dissolved in 500 ml of toluene and 200 ml of absolute ethanol. The solution was cooled to -10°C and HCl(g) was bubbled through until saturation. The mixture was refrigerated for 2 days when most of the solvents were evaporated. Diethyl ether was added and decanted from. The product was redissolved in a solution of O-methylhydroxylamine (10.5 g, 0.125 mol) and triethylamine (56 mL) in 200 mL of methanol. The mixture was allowed to stand for 3 days while the methanol was evaporated with the addition of EtOAc. The organic phase was washed with water, diluted with HOAc and aqueous sodium bicarbonate, dried (Na 2 SO 4 ) and diluted with more EtOAc to a total volume of 500 mL. A 25 ml sample was evaporated to dryness. The remainder was 932 mg. Total yield: 18.6 g (83%).

(ii) 4- aminometvl- N- metoksv- benzamidin (ii) 4-aminomethyl-N-methoxybenzamidine

Til en løsning av 4-azidometyl-N-metoksy-benzamidin (11,3 g, 0,055 mol; se trinn (i) ovenfor) i 200 ml etanol ble tilsatt 200 mg PÆ>2. Blandingen ble hydrogenen med konstant bobling av hydrogen i 4 timer og deretter filtrert gjennom Celite® og inndampet. Utbytte: 7,34 g (74%). To a solution of 4-azidomethyl-N-methoxy-benzamidine (11.3 g, 0.055 mol; see step (i) above) in 200 ml of ethanol was added 200 mg of PÆ>2. The mixture was hydrogenated with constant bubbling of hydrogen for 4 hours and then filtered through Celite® and evaporated. Yield: 7.34 g (74%).

(iii) Boc- Pro- Pab( OMe) (iii) Boc-Pro-Pab(OMe)

Til en suspensjon av Boc-Pro-OH (9,7 g, 0,045 mol), 4-aminometyl-N-metoksy-benzamidin (7,34 g, 0,041 mol; se trinn (ii) ovenfor) og dimetylaminopyridin (7,8 g, 0,064 mol) i 300 ml acetonitril ble tilsatt EDC base (11,7 ml, 0,068 mol). Blandingen ble omrørt i 18 timer, konsentrert og fordelt mellom vann og EtOAc. Det organiske laget ble vasket med vann, vandig natriumbikarbonat, tørket (MgS04) og inndampet. Råproduktet ble flash kromatografert på silikagel med EtOAc. Utbytte: 9,73 g (63%). To a suspension of Boc-Pro-OH (9.7 g, 0.045 mol), 4-aminomethyl-N-methoxy-benzamidine (7.34 g, 0.041 mol; see step (ii) above) and dimethylaminopyridine (7.8 g, 0.064 mol) in 300 ml of acetonitrile was added EDC base (11.7 ml, 0.068 mol). The mixture was stirred for 18 h, concentrated and partitioned between water and EtOAc. The organic layer was washed with water, aqueous sodium bicarbonate, dried (MgSO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc. Yield: 9.73 g (63%).

(iv) H- Pro- Pab( OMe) x 2 HC1 (iv) H-Pro-Pab(OMe) x 2 HCl

Boc-Pro-Pab(OMe) (9,7 g, 0,026 mol; se trinn (iii) ovenfor) ble oppløst i 250 ml EtOAc. Isavkjølt løsning ble mettet med HCl(g) ved bobling i 5 min. Produktet utfelte umiddelbart og 125 ml absolutt etanol ble tilsatt. Blandingen ble ultralydbehandlet inntil mesteparten av materialet hadde stivnet. Dietyleter (200 ml) ble tilsatt og suspensjonen ble filtrert. Noen få klumper som ikke hadde stivnet ble igjen behandlet med absolutt etanol og dietyleter. Det faste stoffet ble tørket. Utbytte: 7,57 g (86%). Boc-Pro-Pab(OMe) (9.7 g, 0.026 mol; see step (iii) above) was dissolved in 250 mL of EtOAc. Ice-cooled solution was saturated with HCl(g) by bubbling for 5 min. The product immediately precipitated and 125 ml absolute ethanol was added. The mixture was ultrasonicated until most of the material had solidified. Diethyl ether (200 mL) was added and the suspension was filtered. A few clumps that had not solidified were again treated with absolute ethanol and diethyl ether. The solid was dried. Yield: 7.57 g (86%).

'H NMR (400 MHz, CD3OD) 8 7,74 (d, 2H), 7,58 (d, 2H), 4,55 (s, 2H), 4,38 (m, 1H), 3,98 (s, 3H), 3,45-3,3 (m, 2H), 2,50 (m, 1H), 2,15-2,0 (m, 3H) 1 H NMR (400 MHz, CD 3 OD) δ 7.74 (d, 2H), 7.58 (d, 2H), 4.55 (s, 2H), 4.38 (m, 1H), 3.98 ( s, 3H), 3.45-3.3 (m, 2H), 2.50 (m, 1H), 2.15-2.0 (m, 3H)

(v) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH)- C( 0)- Pro- Pab( OMe) (v) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH)- C( 0)- Pro- Pab( OMe)

Ph(3-Cl)(5-OCHF2)-WCH(OH)C(0)OH (50 mg, 0,198 mmol; se Eksempel l(viii) ovenfor), H-Pro-Pab(OMe) (76 mg, 0,218 mmol, se trinn (iv) ovenfor) og PyBOP (115 mg, 0,218 mmol) ble oppløst i 2 ml DMF. Diisopropyletylamin (104 mg, 0,80 mmol) ble tilsatt og blandingen ble omrørt i 2,5 timer. Blandingen ble hellet i 50 ml vann og ekstrahert tre ganger med EtOAc og den samlede organiske fase ble vasket med saltvann, tørket (Na2S04) og inndampet. Residuet ble flash kromatografert på silikagel med EtOAc:MeOH (95:5). Utbytte: 37 mg (36%). Renhet: 98%. Ph(3-Cl)(5-OCHF2)-WCH(OH)C(0)OH (50 mg, 0.198 mmol; see Example 1(viii) above), H-Pro-Pab(OMe) (76 mg, 0.218 mmol, see step (iv) above) and PyBOP (115 mg, 0.218 mmol) were dissolved in 2 mL DMF. Diisopropylethylamine (104 mg, 0.80 mmol) was added and the mixture was stirred for 2.5 h. The mixture was poured into 50 mL of water and extracted three times with EtOAc and the combined organic phase was washed with brine, dried (Na 2 SO 4 ) and evaporated. The residue was flash chromatographed on silica gel with EtOAc:MeOH (95:5). Yield: 37 mg (36%). Purity: 98%.

'H NMR (400 MHz, CD3OD blanding av rotamerer) 8 7,60 (d, 2H, hoved rotamer), 7,57 (d, 2H, mindre rotamer), 7,4-7,1 (m, 5H), 6,89 (t, 1H, hoved rotamer), 6,87 (t, 1H, mindre rotamer), 5,35 (s, 1H, hoved rotamer), 5,21 (s, 1H, mindre rotamer), 4,72 (m, 1H, mindre 1H NMR (400 MHz, CD3OD mixture of rotamers) δ 7.60 (d, 2H, major rotamer), 7.57 (d, 2H, minor rotamer), 7.4-7.1 (m, 5H), 6.89 (t, 1H, major rotamer), 6.87 (t, 1H, minor rotamer), 5.35 (s, 1H, major rotamer), 5.21 (s, 1H, minor rotamer), 4, 72 (m, 1H, less

rotamer), 4,5-4,35 (m, 1H og 2H, hoved rotamer), 4,3-4,25 (m, 2H, mindre rotamer), 3,814 (s, 3H, hoved rotamer), 3,807 (s, 3H, mindre rotamer), 3,75-3,5 (m, 1H), 3,35 (m, 1H), 2,2-1,8 (m, 4H) rotamer), 4.5-4.35 (m, 1H and 2H, major rotamer), 4.3-4.25 (m, 2H, minor rotamer), 3.814 (s, 3H, major rotamer), 3.807 (s , 3H, minor rotamer), 3.75-3.5 (m, 1H), 3.35 (m, 1H), 2.2-1.8 (m, 4H)

<l3>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 173,3,173,2,171,3,171,0,153,9,152,4 <l3>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 173,3,173,2,171,3,171,0,153,9,152,4

APCI-MS: (M + 1) = 511/513 m/z APCI-MS: (M + 1) = 511/513 m/z

Eksempel 34 Example 34

Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( 0)- Aze- NH- CH?-(( 2- amidino)- 5- pvridinvl) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( 0)- Aze- NH- CH?-(( 2- amidino)- 5- pvridinvl)

(i) 6- cvanonikotinsvre (i) 6-Cvanonicotin acid

Til en løsning av nikotinsyre N-oksyd (51 g, 0,37 mol) i 1,2 L av DMF, ble NaCN (54 g, 1,1 mol) tilsatt, fulgt av trietylamin (255 ml, 1,83 mol) og TMSC1 (185 ml). Reaksjonsblandingen ble omrørt ved 110°C i 10 timer, filtrert og filtratet ble konsentrert. Residuet ble oppløst i 100 ml 2N HC1 og ekstrahert med metylenklorid. De organiske lag ble samlet, konsentrert og omkrystallisert fra vann, hvilket gir 12 g (22%) av produktet. To a solution of nicotinic acid N-oxide (51 g, 0.37 mol) in 1.2 L of DMF, NaCN (54 g, 1.1 mol) was added, followed by triethylamine (255 mL, 1.83 mol) and TMSC1 (185 ml). The reaction mixture was stirred at 110°C for 10 hours, filtered and the filtrate was concentrated. The residue was dissolved in 100 ml of 2N HCl and extracted with methylene chloride. The organic layers were collected, concentrated and recrystallized from water, yielding 12 g (22%) of the product.

(ii) 5- Qivdroksvmetvl) pvridin- 2- karbonitril (ii) 5- Hydroxymethyl)pyridine-2-carbonitrile

Til en løsning av 6-cyanonikotinsyre (12 g, 0,081 mol; se trinn (i) ovenfor) i THF ved 0°C, Et3N (12,4 ml, 0,0892 mol) ble tilsatt fulgt av etylklorformiat (8,53 ml, 0,0892 mol). Reaksjonsblandingen ble omrørt i 15 min og NaBH} (6,14 g, 0,162 mol) ble tilsatt. Deretter ble blandingen omrørt ved RT natten over, behandlet med vann og ekstrahert med metylenklorid. Det organiske laget ble konsentrert og renset ved kolonnekromatografi, hvilket gir 4 g (20%) av alkoholen. To a solution of 6-cyanonicotinic acid (12 g, 0.081 mol; see step (i) above) in THF at 0 °C, Et 3 N (12.4 mL, 0.0892 mol) was added followed by ethyl chloroformate (8.53 mL , 0.0892 mol). The reaction mixture was stirred for 15 min and NaBH} (6.14 g, 0.162 mol) was added. The mixture was then stirred at RT overnight, treated with water and extracted with methylene chloride. The organic layer was concentrated and purified by column chromatography, yielding 4 g (20%) of the alcohol.

(iii) 5-( Azidometyl) pyridin- 2- karbonitril (iii) 5-(Azidomethyl)pyridine-2-carbonitrile

5-(hydroksymetyl)pyridin-2-karbonitril (4 g, 0,03 mol; se trinn (ii) ovenfor) ble oppløst i 25 ml metylenklorid og avkjølt i et isbad. Mesylklorid (2,32 ml, 0,0300 mol) og deretter trietylamin (4,6 ml, 0,033 mol) ble tilsatt dråpevis. Reaksjonsblandingen ble omrørt og etter opparbeiding ble rå mesylat behandlet med NaN3 (7,35 g, 0,113 mol) i 20 ml DMF. Reaksjonsblandingen ble omrørt ved 40°C i 2 timer, fortynnet med vann og ekstrahert med etylacetat. Det organiske laget ble konsentrert, hvilket gir 3,95 g (83%) av rå azid. 5-(Hydroxymethyl)pyridine-2-carbonitrile (4 g, 0.03 mol; see step (ii) above) was dissolved in 25 mL of methylene chloride and cooled in an ice bath. Mesyl chloride (2.32 mL, 0.0300 mol) and then triethylamine (4.6 mL, 0.033 mol) were added dropwise. The reaction mixture was stirred and after workup the crude mesylate was treated with NaN 3 (7.35 g, 0.113 mol) in 20 mL DMF. The reaction mixture was stirred at 40°C for 2 hours, diluted with water and extracted with ethyl acetate. The organic layer was concentrated to give 3.95 g (83%) of crude azide.

(iv) 5-( ferr- butoksvkarbonylaminometvl) pvridin- 2- karbonitril (iv) 5-(ferr-butoxycarbonylaminomethyl)pyridine-2-carbonitrile

Til en løsning av 5-(azidometyl)pyridin-2-karbonitril (3,95 g, 0,0248 mol; se trinn (iii) ovenfor) i 30 ml THF og 10 ml vann, trifenylfosfin (7,8 g, 0,0298 mol) ble tilsatt og det resulterende omrørt i 24 timer. Deretter, ble trietylamin (3,8 ml, 0,027 mol) tilsatt, fulgt av Boc-anhydrid (5,4 g, 0,025 mol) og omrøring i 2 timer. Reaksjonsblandingen ble fordelt mellom vann og etylacetat. Det organiske laget ble konsentrert og renset ved To a solution of 5-(azidomethyl)pyridine-2-carbonitrile (3.95 g, 0.0248 mol; see step (iii) above) in 30 mL THF and 10 mL water, triphenylphosphine (7.8 g, 0, 0298 mol) was added and the resulting stirred for 24 hours. Then, triethylamine (3.8 mL, 0.027 mol) was added, followed by Boc anhydride (5.4 g, 0.025 mol) and stirring for 2 h. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was concentrated and purified by

. kolonnekromatografi, hvilket gir 2,1 g (36%) av sub-tittelforbindelsen. . column chromatography, yielding 2.1 g (36%) of the sub-title compound.

'H NMR (300 MHz, CDC13) 8 8,6 (s, 1H), 8,0 (d, 1H), 8,9 (d, 1H), 4,1 (m, 2H), 1,4 (s, 9H) 1 H NMR (300 MHz, CDCl 3 ) δ 8.6 (s, 1H), 8.0 (d, 1H), 8.9 (d, 1H), 4.1 (m, 2H), 1.4 ( pp. 9H)

(v) 5-( aminometvl) pvridin- 2- karbonitril x 2 HC1 (v) 5-(aminomethyl)pyridine-2-carbonitrile x 2 HCl

5-(teAt-butoksykarbonylaminometyl)pyridin-2-karbonitril (0,200 g, 0,86 mmol, se trinn (iv) ovenfor) ble oppløst i 10 ml EtOAc mettet med HCl(g) og ble omrørt i 30 min. Løsningsmidlet ble avdampet og 0,175 g (99%) av sub-tittelforbindelse ble oppnådd som dens dihydrokloridsalt. 5-(teAt-butoxycarbonylaminomethyl)pyridine-2-carbonitrile (0.200 g, 0.86 mmol, see step (iv) above) was dissolved in 10 mL EtOAc saturated with HCl(g) and stirred for 30 min. The solvent was evaporated and 0.175 g (99%) of the sub-title compound was obtained as its dihydrochloride salt.

'H NMR (500 MHz, D20) 8 8,79 (s, 1H), 8,17 (d, 1H), 8,05 (d, 1H), 4,38 (s, 2H) 1 H NMR (500 MHz, D 2 O) δ 8.79 (s, 1H), 8.17 (d, 1H), 8.05 (d, 1H), 4.38 (s, 2H)

(vi) Boc- Aze- NH- CH9- 5- Pv( 2- CN) (vi) Boc- Aze- NH- CH9- 5- Pv( 2- CN)

Til en blanding av 5-(aminometyl)pyridin-2-karbonitril x 2 HC1 (0,175 g, 0j85 mmol; se trinn (v) ovenfor), Boc-Aze-OH (0,201 g, 1,00 mmol) og TBTU (0,321 g, 1,00 mmol) i 5 ml DMF ble det tilsatt dimetylaminopyridin (0,367 g, 3,00 mmol). Blandingen ble omrørt natten over og deretter hellet i vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vandig natriumbikarbonat, tørket (Na2S04) og inndampet. Råproduktet begynte å krystallisere og ble anvendt som sådanne i neste trinn. Utbytte: 0,23 g (73%). To a mixture of 5-(aminomethyl)pyridine-2-carbonitrile x 2 HCl (0.175 g, 0.85 mmol; see step (v) above), Boc-Aze-OH (0.201 g, 1.00 mmol) and TBTU (0.321 g, 1.00 mmol) in 5 ml DMF was added dimethylaminopyridine (0.367 g, 3.00 mmol). The mixture was stirred overnight and then poured into water and extracted three times with EtOAc. The combined organic phase was washed with aqueous sodium bicarbonate, dried (Na 2 SO 4 ) and evaporated. The crude product began to crystallize and was used as such in the next step. Yield: 0.23 g (73%).

'H NMR (500 MHz, CDC13) 8 8,66 (s, 1H), 8,2-7,8 (bred, 1H), 7,79 (d, 1H), 7,67 (d, 1H), 4,73 (m, 1H), 4,65-4,5 (m, 2H), 3,94 (m, 1H), 3,81 (m, 1H), 2,6,2,35 (m, 2H), 1,8 (bred, 1H), 1,45 (s, 9H) 1 H NMR (500 MHz, CDCl 3 ) δ 8.66 (s, 1H), 8.2-7.8 (broad, 1H), 7.79 (d, 1H), 7.67 (d, 1H), 4.73 (m, 1H), 4.65-4.5 (m, 2H), 3.94 (m, 1H), 3.81 (m, 1H), 2.6, 2.35 (m, 2H), 1.8 (wide, 1H), 1.45 (s, 9H)

(vii) H- Aze- NH- CHr5- Pv( 2- CN) x 2 HC1 (vii) H-Aze-NH-CHr5-Pv(2-CN) x 2 HCl

Boc-Aze-NH-CH2-5-Py(2-CN) (0,23 g, 0,73 mmol; se trinn (vi) ovenfor) ble oppløst i 10 ml EtOAc mettet med HCl(g) og ble omrørt i 30 min. Løsningsmidlet ble avdampet og 0,21 g (100%) av sub-tittelforbindelsen ble oppnådd som dens dihydrokloridsalt. Boc-Aze-NH-CH2-5-Py(2-CN) (0.23 g, 0.73 mmol; see step (vi) above) was dissolved in 10 mL of EtOAc saturated with HCl(g) and was stirred in 30 min. The solvent was evaporated and 0.21 g (100%) of the sub-title compound was obtained as its dihydrochloride salt.

<*>H NMR (500 MHz, D20) 8 8,64 (s, 1H), 8,0-7,9 (m, 2H), 5,19 (m, 1H), 4,65-4,55 (m, 2H), 4,20 (m, 1H), 4,03 (m, 1H), 2,88 (m, 1H), 2,64 (m, 1H) <*>H NMR (500 MHz, D 2 O) δ 8.64 (s, 1H), 8.0-7.9 (m, 2H), 5.19 (m, 1H), 4.65-4.55 (m, 2H), 4.20 (m, 1H), 4.03 (m, 1H), 2.88 (m, 1H), 2.64 (m, 1H)

(viii) Ph( 3- Cl)( 5- OCHFz)-(/ ?JCH( OH)- C( Q)- Aze- NH- CH2- 5- Pv( 2- CN) (viii) Ph( 3- Cl)( 5- OCHFz)-(/ ?JCH( OH)- C( Q)- Aze- NH- CH2- 5- Pv( 2- CN)

Til en blanding av H-Aze-NH-CH2-5-Py(2-CN) x 2 HC1 (0,206 g, 0,713 mmol; se trinn (vii) ovenfor), Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)OH (0,180 g, 0,713 mmol; se Eksempel l(viii) ovenfor) og PyBOP (0,408 g, 0,784 mmol) i 5 ml DMF ble tilsatt dimetylaminopyridin (0,367 g, 3,00 mmol). Blandingen ble omrørt natten over og deretter hellet i vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vandig natriumbikarbonat, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel med EtOAc og ga et rent produkt. Utbytte: 0,197 g (61%). To a mixture of H-Aze-NH-CH2-5-Py(2-CN) x 2 HCl (0.206 g, 0.713 mmol; see step (vii) above), Ph(3-Cl)(5-OCHF2)- (/?>)CH(OH)C(0)OH (0.180 g, 0.713 mmol; see Example 1(viii) above) and PyBOP (0.408 g, 0.784 mmol) in 5 mL DMF was added dimethylaminopyridine (0.367 g, 3 .00 mmol). The mixture was stirred overnight and then poured into water and extracted three times with EtOAc. The combined organic phase was washed with aqueous sodium bicarbonate, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc to give a pure product. Yield: 0.197 g (61%).

'H NMR (500 MHz, CDC13) 8 8,63 (m, 1H), 8,22 (bt, 1H), 7,78 (m, 1H), 7,67 (m, 1H), 7,21 (m, 1H), 7,16 (m, 1H), 7,04 (m, 1H), 6,56 (t, 1H), 4,97 (bd, 1H), 4,92 (m, 1H), 4,6-4,5 (m, 2H), 4,40 (bd, 1H), 4,18 (m, 1H), 3,80 (m, 1H), 2,69 (m, 1H), 2,46 (m, 1H), 1,92 (s, 1H) 1 H NMR (500 MHz, CDCl 3 ) δ 8.63 (m, 1H), 8.22 (bt, 1H), 7.78 (m, 1H), 7.67 (m, 1H), 7.21 ( m, 1H), 7.16 (m, 1H), 7.04 (m, 1H), 6.56 (t, 1H), 4.97 (bd, 1H), 4.92 (m, 1H), 4.6-4.5 (m, 2H), 4.40 (bd, 1H), 4.18 (m, 1H), 3.80 (m, 1H), 2.69 (m, 1H), 2 .46 (m, 1H), 1.92 (s, 1H)

APCI-MS: (M + 1) = 451/453 m/z APCI-MS: (M + 1) = 451/453 m/z

(ix) Ph( 3- Cl)( 5- OCHF9)-(/ g) CH( OH)- C( Q)- Aze- NH- CH7-(( 2- amidino)- 5- (ix) Ph( 3- Cl)( 5- OCHF9)-(/ g) CH( OH)- C( Q)- Aze- NH- CH7-(( 2- amidino)- 5-

p<y>ridin<y>l) x HOAc p<y>ridin<y>l) x HOAc

Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)-C(0)-Aze-NH-CH2-5-Py(2-CN) (0,200 g, 0,444 mmol; se trinn (viii) ovenfor), ammoniumacetat (1,00 g, 0,0130 mol) og N-acetylcystein (2,00 g, 0,0122 mol) i 10 ml metanol ble oppvarmet ved 50°C i 2 dager. Preparativ RPLC med CH3CN:0,1M NH4OAC (30:79) og kjøring av de passende fraksjoner igjen med CH3CN:0,1M NHjOAc (5:95 - 40:60) ga 60 mg (26%) av ren tittelforbindelse som dens acetatsalt etter fryse tørking fra vann og acetonitril. Renhet: 100%. Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)-C(0)-Aze-NH-CH2-5-Py(2-CN) (0.200 g, 0.444 mmol; see step (viii) above), ammonium acetate (1.00 g, 0.0130 mol) and N-acetylcysteine (2.00 g, 0.0122 mol) in 10 mL methanol was heated at 50°C for 2 days. Preparative RPLC with CH3CN:0.1M NH4OAC (30:79) and running the appropriate fractions again with CH3CN:0.1M NHjOAc (5:95 - 40:60) gave 60 mg (26%) of pure title compound as its acetate salt after freeze drying from water and acetonitrile. Purity: 100%.

'H NMR (500 MHz, D20, blanding av rotamerer) 8 8,68 (s, 1H, hoved rotamer), 8,62 (s, 1H, mindre rotamer), 8,05-7,9 (m, 2H), 7,33 (m, 1H, rotamer), 7,27 (m, 1H, rotamer), 7,22 (m, 1H, rotamer), 7,17 (m, 1H, rotamer), 7,01 (m, 1H, rotamer), 6,84 (t, 1H), 5,32 (s, 1H, hoved rotamer), 5,20 (m, 1H, mindre rotamer), 5,13 (s, 1H, mindre rotamer), 4,88 (m, 1H, hoved rotamer), 4,65-4,55 (m, 2H, hoved rotamer), 4,45-4,35 (m, 1H, rotamer pluss 1H, mindre rotamer), 4,31 (d, 1H, mindre rotamer), 4,2-4,05 (m, 1H pluss 1H, rotamer), 2,80 (m, 1H, mindre rotamer), 2,61 (m, 1H, hoved rotamer), 2,33 (m, 1H, hoved rotamer), 2,24 (m, 1H, mindre rotamer), 1,93 (s, 3H) 1H NMR (500 MHz, D 2 O, mixture of rotamers) δ 8.68 (s, 1H, major rotamer), 8.62 (s, 1H, minor rotamer), 8.05-7.9 (m, 2H) , 7.33 (m, 1H, rotamer), 7.27 (m, 1H, rotamer), 7.22 (m, 1H, rotamer), 7.17 (m, 1H, rotamer), 7.01 (m , 1H, rotamer), 6.84 (t, 1H), 5.32 (s, 1H, major rotamer), 5.20 (m, 1H, minor rotamer), 5.13 (s, 1H, minor rotamer) , 4.88 (m, 1H, major rotamer), 4.65-4.55 (m, 2H, major rotamer), 4.45-4.35 (m, 1H, rotamer plus 1H, minor rotamer), 4 .31 (d, 1H, minor rotamer), 4.2-4.05 (m, 1H plus 1H, rotamer), 2.80 (m, 1H, minor rotamer), 2.61 (m, 1H, major rotamer ), 2.33 (m, 1H, major rotamer), 2.24 (m, 1H, minor rotamer), 1.93 (s, 3H)

C-NMR (100 MHz; D20): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 181,6,173,3,172,7,172,6,172,3,162,6,162,3 C-NMR (100 MHz; D20): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 181,6,173,3,172,7,172,6,172,3,162,6,162,3

APCI-MS: (M + 1) = 468/470 m/z APCI-MS: (M + 1) = 468/470 m/z

Eksempel 35 Example 35

Ph( 3- Cl)( 5- OCHF9)-(/ g) CH( OH) C( 0)- Aze- NH- CHr(( 2- metoksvamidino)- 5- pvridinvl) Ph( 3- Cl)( 5- OCHF9)-(/ g) CH( OH) C( 0)- Aze- NH- CHr(( 2- methoxyvamidino)- 5- pvridinvl)

(i) Boc- NH- CH7- r( 2-( aminoQivdroksvlimino) metvl))- 5- pvridinvll 5-(te/t-butoksykarbonylaminometyl)pyridin-2-karbonitril (1,00 g, 4,29 mmol; se Eksempel 34(iv) ovenfor) ble oppløst i 10 ml etanol og hydroksylamin-hydroklorid (0,894 g, 0,0129 mol) og trietylamin (1,30 g, 0,0129 mol) ble tilsatt. Blandingen ble omrørt ved romtemperatur i 6 dager. Blandingen ble fordelt mellom vann og metylenklorid. Det vandige laget ble ekstrahert med metylenklorid og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Utbytte: 0,96 g (84%). (i) Boc-NH-CH7-r(2-(aminohydroxylimino)methyl)-5-pyridinyl 5-(t-butoxycarbonylaminomethyl)pyridine-2-carbonitrile (1.00 g, 4.29 mmol; see Example 34(iv) above) was dissolved in 10 ml of ethanol and hydroxylamine hydrochloride (0.894 g, 0.0129 mol) and triethylamine (1.30 g, 0.0129 mol) were added. The mixture was stirred at room temperature for 6 days. The mixture was partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 0.96 g (84%).

<*>H NMR (400 MHz, aceton-de) 8 9,01 (bs, 1H), 8,50 (bs, 1H), 7,87 (m, 1H), 7,70 (m, 1H), 6,58 (bred, 1H), 5,70 (bred, 2H), 4,31 (d, 2H), 1,41 (s, 9H) <*>H NMR (400 MHz, acetone-de) δ 9.01 (bs, 1H), 8.50 (bs, 1H), 7.87 (m, 1H), 7.70 (m, 1H), 6.58 (wide, 1H), 5.70 (wide, 2H), 4.31 (d, 2H), 1.41 (s, 9H)

(ii) Boc- Aze- NH- CH2-( 2-( amidino)- 5- pvridinvn x HOAc (ii) Boc- Aze- NH- CH2-( 2-( amidino)- 5- pvridinvn x HOAc

Denne reaksjonen ble utført i henhold til metoden beskrevet i Judkins et al, Synth. Comm. This reaction was carried out according to the method described in Judkins et al, Synth. Comm.

(1998) 4351. En suspensjon av Boc-NH-CH2-[(2-(amino(hydroksylimino)metyl))-5-pyridinyl] (0,910 g, 3,42 mmol; se trinn (i) ovenfor), eddiksyreanhydrid (0,35 ml, 3,7 mmol) og 0,35 g 10% Pd/C (50% fuktighet) i 100 ml eddiksyre ble hydrogenen ved et trykk på 5 atm. i 5 timer. Blandingen ble filtrert gjennom Celite og konsentrert. Residuet ble frysetørket fra vann og acetonitril, hvilket gir 0,97 g (92%) av sub-tittelforbindelse. (1998) 4351. A suspension of Boc-NH-CH2-[(2-(amino(hydroxylimino)methyl))-5-pyridinyl] (0.910 g, 3.42 mmol; see step (i) above), acetic anhydride ( 0.35 ml, 3.7 mmol) and 0.35 g of 10% Pd/C (50% moisture) in 100 ml of acetic acid became the hydrogen at a pressure of 5 atm. for 5 hours. The mixture was filtered through Celite and concentrated. The residue was freeze-dried from water and acetonitrile to give 0.97 g (92%) of sub-title compound.

•h NMR (500 MHz, CD3OD) 8 8,74 (s, 1H), 8,12 (d, 1H), 7,98 (d, 1H), 4,38 (s, 2H), 1,92 (s, 3H),1,46 (s,9H) •h NMR (500 MHz, CD3OD) δ 8.74 (s, 1H), 8.12 (d, 1H), 7.98 (d, 1H), 4.38 (s, 2H), 1.92 ( p, 3H),1.46 (p,9H)

(iii) Boc- NH- CH2-( 2-( amino( trimetvlsilyletvlimino) metvl)- 5- pvridinvl) (iii) Boc-NH-CH2-(2-(amino(trimethylsilylethlimino)methyl)-5- pvridinyl)

Til en suspensjon av Boc-NH-CH2-(2-(amidino)-5-pyridinyl) x HOAc (0,96 g, 3,1 mmol; se trinn (ii) ovenfor) i 75 ml THF ble det tilsatt en løsning av kaliumkarbonat (1,07 g, 7,7 mmol) og Teoc-/7-nitrofenylkarbonat (1,14 g, 4,02 mmol) i 15 ml vann. Blandingen ble omrørt natten over. Et overskudd av glycin og kaliumkarbonat ble tilsatt og reaksjonen ble fortsatt i 2 timer. THF ble inndampet og resten ble ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Produktet kunne anvendes uten ytterligere rensning. To a suspension of Boc-NH-CH2-(2-(amidino)-5-pyridinyl) x HOAc (0.96 g, 3.1 mmol; see step (ii) above) in 75 mL of THF was added a solution of potassium carbonate (1.07 g, 7.7 mmol) and Teoc-β-nitrophenyl carbonate (1.14 g, 4.02 mmol) in 15 mL of water. The mixture was stirred overnight. An excess of glycine and potassium carbonate was added and the reaction was continued for 2 hours. The THF was evaporated and the residue was extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification.

'H NMR (500 MHz, CDC13) 8 9,31 (bred, 1H), 8,52 (s, 1H), 8,41 (d, 1H), 8,35 (bred, 1H), 7,74 (d, 1H), 4,97 (bred, 1H), 4,39 (m, 2H), 4,26 (m, 2H), 1,46 (s, 9H), 1,14 (m, 2H), 0,07 1 H NMR (500 MHz, CDCl 3 ) δ 9.31 (broad, 1H), 8.52 (s, 1H), 8.41 (d, 1H), 8.35 (broad, 1H), 7.74 ( d, 1H), 4.97 (b, 1H), 4.39 (m, 2H), 4.26 (m, 2H), 1.46 (s, 9H), 1.14 (m, 2H), 0.07

(s, 9H) (pp, 9H)

(iv) H7N- CH2-( 2-( amino( trimetylsilvletvlimino) metvl)- 5- pvridinyl) x 2 HC1 Boc-NH-CH2-(2-(amino(trimetylsilyletylimino)metyl)-5-pyridinyl) (0,23 g, 0,58 mmol; se trinn (iii) ovenfor) ble oppløst i 25 ml EtOAc mettet med HCl(g) og omrørt i 30 min. Løsningsmidlet ble avdampet og produktet anvendt uten ytterligere rensning. Utbytte: 0,21 g (98%). (iv) H7N- CH2-( 2-(amino(trimethylsilylethylimino)methyl)-5- pvridinyl) x 2 HC1 Boc-NH-CH2-(2-(amino(trimethylsilylethylimino)methyl)-5-pyridinyl) (0.23 g, 0.58 mmol; see step (iii) above) was dissolved in 25 mL of EtOAc saturated with HCl(g) and stirred for 30 min. The solvent was evaporated and the product used without further purification. Yield: 0.21 g (98%).

'H NMR (500 MHz, D20) 5 8,89 (s, 1H), 8,25 (s, 2H), 4,55 (m, 2H), 4,42 (s, 2H), 1,20 (m, 2H), 0,09 (s, 9H) 1 H NMR (500 MHz, D 2 O) δ 8.89 (s, 1H), 8.25 (s, 2H), 4.55 (m, 2H), 4.42 (s, 2H), 1.20 ( m, 2H), 0.09 (s, 9H)

(v) Boc- Aze- NH- CH9-( 2-( amino( trimetvlsilvletvlimino) metvl)- 5- (v) Boc- Aze- NH- CH9-( 2-( amino(trimethylsilvletylimino) methyl)- 5-

pvridinvl) pvridinvl)

Til en løsning av H2N-CH2-(2-(amino(trimetylsilyletylimino)metyl)-5-pyridinyl) x 2 HC1 (0,21 g, 0,57 mmol; se trinn (iv) ovenfor), Boc-Aze-OH (0,127 g, 0,631 mmol) og TBTU (233 mg, 0,726 mmol) i 5 ml DMF ble tilsatt dimetylaminopyridin (269 mg, 2,20 mmol). Blandingen ble omrørt natten over, hellet i 100 ml vann og ekstrahert med EtOAc tre ganger. Den samlede organiske fase ble vasket med vandig natriumbikarbonat og vann, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel med EtOAc, hvilket gir 170 mg (56%) av det ønskede produkt. To a solution of H2N-CH2-(2-(amino(trimethylsilylethylimino)methyl)-5-pyridinyl) x 2 HCl (0.21 g, 0.57 mmol; see step (iv) above), Boc-Aze-OH (0.127 g, 0.631 mmol) and TBTU (233 mg, 0.726 mmol) in 5 mL DMF was added dimethylaminopyridine (269 mg, 2.20 mmol). The mixture was stirred overnight, poured into 100 mL water and extracted with EtOAc three times. The combined organic phase was washed with aqueous sodium bicarbonate and water, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc, which gives 170 mg (56%) of the desired product.

'H NMR (500 MHz, CDC13) 6 9,33 (bred, 1H), 8,54 (s, 1H), 8,41 (d, 1H), 8,36 (bred, 1H), 7,75 (m, 1H), 4,72 (m, 1H), 4,56 (m, 2H), 4,26 (m, 2H), 3,93 (m, 1H), 3,80 (m, 1H), 2,6-2,4 (m, 2H), 1,42 (s, 9H), 1,14 (m, 2H), 0,07 (s, 9H) 1 H NMR (500 MHz, CDCl 3 ) δ 9.33 (broad, 1H), 8.54 (s, 1H), 8.41 (d, 1H), 8.36 (broad, 1H), 7.75 ( m, 1H), 4.72 (m, 1H), 4.56 (m, 2H), 4.26 (m, 2H), 3.93 (m, 1H), 3.80 (m, 1H), 2.6-2.4 (m, 2H), 1.42 (s, 9H), 1.14 (m, 2H), 0.07 (s, 9H)

(vi) H- Aze- NH- CH2-( 2-( amino( trimetvlsilvletvlimino) metvl)- 5- (vi) H- Aze- NH- CH2-( 2-( amino(trimethylsilvletylimino) methyl)- 5-

pvridinvl) x 2 HC1 pvridinvl) x 2 HCl

Boc-Aze-NH-CH2-(2-(amino(trimetylsilyletylimino)metyl)-5-pyridinyl) (170 mg, 0,356 mmol; se trinn (v) ovenfor) ble oppløst i 25 ml EtOAc mettet med HCl(g) og omrørt i 30 min. Løsningsmidlet ble avdampet og produktet anvendt uten ytterligere rensning. Utbytte: 160 mg (100%). Boc-Aze-NH-CH2-(2-(amino(trimethylsilylethylimino)methyl)-5-pyridinyl) (170 mg, 0.356 mmol; see step (v) above) was dissolved in 25 mL of EtOAc saturated with HCl(g) and stirred for 30 min. The solvent was evaporated and the product used without further purification. Yield: 160 mg (100%).

<J>H NMR (500 MHz, CD3OD) 8 9,00 (m, 1H), 8,84 (m, 1H), 8,23 (d, 2H), 8,10 (m, 1H), 5,09 (m, 1H), 4,7-4,6 (m, 2H), 4,51 (m, 2H), 4,14 (m, 1H), 3,97 (m, 1H), 2,86 (m, 1H), 2,58 (m, 1H), 1,22 (m, 2H), 0,11 (s, 9H) <J>H NMR (500 MHz, CD3OD) δ 9.00 (m, 1H), 8.84 (m, 1H), 8.23 (d, 2H), 8.10 (m, 1H), 5, 09 (m, 1H), 4.7-4.6 (m, 2H), 4.51 (m, 2H), 4.14 (m, 1H), 3.97 (m, 1H), 2.86 (m, 1H), 2.58 (m, 1H), 1.22 (m, 2H), 0.11 (s, 9H)

(vii) Ph( 3- Cl)( 5- OCHF7)- r/ ?) CH( OH) C( 0)- Aze- NH- CHz-( 2-( amino( tri-metvlsilvletylimino) metvl)- 5- pvridinvl) (vii) Ph( 3- Cl)( 5- OCHF7)- r/ ?) CH( OH) C( 0)- Aze- NH- CHz-( 2-( amino( tri-methylsilvethylimino) methyl)- 5- pvridinvl )

Til en løsning av H-Aze-NH-CH2-(2-(amino(trimetylsilyletylimino)-metyl)-5-pyridinyl) x 2 HC1 (160 mg, 0,462 mmol; se trinn (vi) ovenfor), Ph(3-Cl)(5-OCHF2)-r/?;CH(OH)C(0)OH (131 mg, 0,462 mmol; se Eksempel l(viii) ovenfor) og PyBOP (263 mg, 0,505 mmol) i 5 ml DMF ble det tilsatt diisopropyletylamin (0,30 ml, 1,71 mmol). Blandingen ble omrørt natten over, hellet i 100 ml vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vandig natriumbikarbonat og vann, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel med EtOAc:MeOH (95:5), hvilket gir 148 mg (52%) av det ønskede produkt. To a solution of H-Aze-NH-CH2-(2-(amino(trimethylsilylethylimino)-methyl)-5-pyridinyl) x 2 HCl (160 mg, 0.462 mmol; see step (vi) above), Ph(3- Cl)(5-OCHF2)-r/?;CH(OH)C(0)OH (131 mg, 0.462 mmol; see Example 1(viii) above) and PyBOP (263 mg, 0.505 mmol) in 5 mL DMF were diisopropylethylamine (0.30 mL, 1.71 mmol) was added. The mixture was stirred overnight, poured into 100 mL water and extracted three times with EtOAc. The combined organic phase was washed with aqueous sodium bicarbonate and water, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with EtOAc:MeOH (95:5), which gives 148 mg (52%) of the desired product.

(viii) Ph( 3- Cl)( 5- OCHF2)- ri?) CH( OH) C( 0)- Aze- NH- CH?-( 2-( metoksv-amino( trimetvlsilvletvlimino) metvl)- 5- pvridinyl) (viii) Ph( 3- Cl)( 5- OCHF 2 )- ri?) CH( OH) C( 0)- Aze- NH- CH?-( 2-( methoxys-amino(trimethylsylsilvletvlimino) methyl)- 5- pyridinyl )

En suspensjon av Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)-C(0)-Aze-NH-CH2-(2-(metoksyamino(trimetylsilyletylimino)metyl)-5-pyridinyl) (148 mg, 0,242 mmol; se trinn (vii) ovenfor) og O-metylhydroksyl amin (202 mg, 2,42 mmol) i 10 ml acetonitril ble oppvarmet ved 70°C i 3 timer. Blandingen ble fordelt mellom vann og EtOAc. Det vandige laget ble ekstrahert to ganger med EtOAc og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Rå materialet ble flash kromatografert på silikagel med EtOAc:MeOH (95:5), hvilket gir 44 mg (28%) av rent materiale. A suspension of Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)-C(0)-Aze-NH-CH2-(2-(methoxyamino(trimethylsilylethylimino)methyl)-5-pyridinyl) (148 mg, 0.242 mmol; see step (vii) above) and O-methylhydroxyl amine (202 mg, 2.42 mmol) in 10 mL of acetonitrile was heated at 70 °C for 3 h. The mixture was partitioned between water and EtOAc. The aqueous layer was extracted twice with EtOAc and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The crude material was flash chromatographed on silica gel with EtOAc:MeOH (95:5), giving 44 mg (28%) of pure material.

'H NMR (500 MHz, CDC13) 8 8,55 (m, 1H), 8,05 (bt, 1H), 7,70 (m, 1H), 7,58 (s, 1H), 7,56 (d, 1H), 7,22 (m, 1H), 7,16 (m, 1H), 7,03 (m, 1H), 6,50 (t, 1H), 4,92 (s, 1H), 4,89 (m, 1H), 4,55-4,45 (m, 2H), 4,38 (bred, 1H), 4,2-4,1 (m, 3H), 4,00 (s, 3H), 3,73 (m, 1H), 2,69 (m, 1H), 2,44 (m, 1H), 0,97 (m, 2H), 0,02 (s, 9H) 1 H NMR (500 MHz, CDCl 3 ) δ 8.55 (m, 1H), 8.05 (bt, 1H), 7.70 (m, 1H), 7.58 (s, 1H), 7.56 ( d, 1H), 7.22 (m, 1H), 7.16 (m, 1H), 7.03 (m, 1H), 6.50 (t, 1H), 4.92 (s, 1H), 4.89 (m, 1H), 4.55-4.45 (m, 2H), 4.38 (wide, 1H), 4.2-4.1 (m, 3H), 4.00 (s, 3H), 3.73 (m, 1H), 2.69 (m, 1H), 2.44 (m, 1H), 0.97 (m, 2H), 0.02 (s, 9H)

(ix) Ph( 3- Cl)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- NH- CH2-(( 2- metoksv-amidino)- 5- pvridinvl) (ix) Ph( 3- Cl)( 5- OCHF?)-(/ g) CH( OH) C( 0)- Aze- NH- CH2-(( 2- methoxysv-amidino)- 5- pvridinvl)

Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-NH-CH2-(2-(metoksyamino(tri-metylsilyletylimino)metyl)-5-pyridinyl) (44 mg, 0,069 mmol; se trinn (viii) ovenfor) ble oppløst i 2 ml TFA og fikk reagere i 1 time. TFA ble inndampet og residuet ble fordelt mellom EtOAc og vandig natriumbikarbonat. Det vandige laget ble ekstrahert med EtOAc og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Utbytte: 30 mg (88%). Renhet: >95%. Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-NH-CH2-(2-(methoxyamino(tri-methylsilylethylimino)methyl)-5-pyridinyl) ( 44 mg, 0.069 mmol; see step (viii) above) was dissolved in 2 mL of TFA and allowed to react for 1 hour. The TFA was evaporated and the residue was partitioned between EtOAc and aqueous sodium bicarbonate. The aqueous layer was extracted with EtOAc and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 30 mg (88%). Purity: >95%.

'H NMR (500 MHz, CDC13) 8 8,44 (m, 1H), 8,03 (bt, 1H), 7,91 (m, 1H), 7,60 (m, 1H), 7,19 (m, 1H), 7,13 (m, 1H), 7,00 (m, 1H), 6,52 (t, 1H), 5,6-5,45 (bred, 2H), 4,90 (s, 1H), 4,89 (m, 1H), 4,55-4,4 (m, 2H), 4,27 (bred, 1H), 4,12 (m, 1H), 3,92 (s, 3 H), 2,68 (m, 1H), 2,41 (m, 1H) 1 H NMR (500 MHz, CDCl 3 ) δ 8.44 (m, 1H), 8.03 (bt, 1H), 7.91 (m, 1H), 7.60 (m, 1H), 7.19 ( m, 1H), 7.13 (m, 1H), 7.00 (m, 1H), 6.52 (t, 1H), 5.6-5.45 (wide, 2H), 4.90 (s , 1H), 4.89 (m, 1H), 4.55-4.4 (m, 2H), 4.27 (broad, 1H), 4.12 (m, 1H), 3.92 (s, 3H), 2.68 (m, 1H), 2.41 (m, 1H)

<13>C-NMR (100 MHz; CDC13): (karbonyl og/eller amidinkarbonatomer) 8 173,0,170,9, 152,6 <13>C-NMR (100 MHz; CDC13): (carbonyl and/or amidine carbon atoms) 8 173.0, 170.9, 152.6

APCI-MS: (M + 1) = 498/500 m/z APCI-MS: (M + 1) = 498/500 m/z

Eksempel 36 Example 36

Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( 0)- Aze- NH- CH7-(( 5- amidino)- 2- pvrimidinvl) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( 0)- Aze- NH- CH7-(( 5- amidino)- 2- pvrimidinvl)

(i) 2- amino- 2- iminoetvlkarbamafAcOH (i) 2-amino-2-iminoethylcarbamafAcOH

N-Boc-aminoacetonitril (40,2 g, 257,4 mmol) og N-acetylcystein (42,0 g, 257,4 mmol) ble oppløst i metanol (300 ml) ved 60°C og ammoniakk ble ført gjennom i 18 timer. Løsningsmidlet ble fjernet i vakuum. Etter ionebytter kromatografi (Amberlite IRA-400 (AcOH)) og omkrystallisering fra aceton, ble 28,4 g (53%) av sub-tittelforbindelsen oppnådd som et hvitt, fast stoff. N-Boc-aminoacetonitrile (40.2 g, 257.4 mmol) and N-acetylcysteine (42.0 g, 257.4 mmol) were dissolved in methanol (300 mL) at 60°C and ammonia was passed through for 18 hours. The solvent was removed in vacuo. After ion exchange chromatography (Amberlite IRA-400 (AcOH)) and recrystallization from acetone, 28.4 g (53%) of the sub-title compound was obtained as a white solid.

'H NMR (300 MHz, CD3OD) 8 4,41 (t, J = 4,9 Hz, 1H), 4,01 (s, 2H), 2,91 (d, J = 5,0 Hz, 2H), 2,01 (s, 3H), 1,46 (s, 9H) 1H NMR (300 MHz, CD3OD) δ 4.41 (t, J = 4.9 Hz, 1H), 4.01 (s, 2H), 2.91 (d, J = 5.0 Hz, 2H) , 2.01 (s, 3H), 1.46 (s, 9H)

(ii) 1. 3- bis( dimetvlamino)- 2- cvanotrimethiniumperklorat (ii) 1. 3- bis(dimethylamino)- 2- cvanotrimethinium perchlorate

En løsning av 3-dimetylaminoakrylonitril (25,0 g, 260,0 mmol) i kloroform (75 ml) ble satt dråpevis til en løsning av (klormetylen)dimetylammoniumklorid (50,0 g, 390,1 mmol) i kloroform (175 ml) ved 0°C. Reaksjonsblandingen ble omrørt i ytterligere 2 timer ved 0°C, og fikk deretter oppvarmes til romtemperatur natten over, deretter oppvarmet i 8 timer under tilbakeløp. Løsningsmidlet ble fjernet i vakuum. Residuet ble satt til en blanding av natriumperklorat (110 g, 0,898 mmol) i vann (150 ml) og etanol (300 ml). Blandingen ble oppvarmet under tilbakeløp i 15 min deretter avkjølt og fikk stå natten over i et kjøleskap. Fellingen ble oppsamlet og omkrystallisert fra etanol, hvilket gir 23,8 g (52%) av sub-tittelforbindelse som fargeløse nåler. A solution of 3-dimethylaminoacrylonitrile (25.0 g, 260.0 mmol) in chloroform (75 mL) was added dropwise to a solution of (chloromethylene)dimethylammonium chloride (50.0 g, 390.1 mmol) in chloroform (175 mL ) at 0°C. The reaction mixture was stirred for an additional 2 hours at 0°C, and then allowed to warm to room temperature overnight, then heated for 8 hours under reflux. The solvent was removed in vacuo. The residue was added to a mixture of sodium perchlorate (110 g, 0.898 mmol) in water (150 mL) and ethanol (300 mL). The mixture was heated under reflux for 15 min then cooled and allowed to stand overnight in a refrigerator. The precipitate was collected and recrystallized from ethanol to give 23.8 g (52%) of the sub-title compound as colorless needles.

Sm.p.: 140-141°C Melting point: 140-141°C

'H NMR (300 MHz, CDC13) 8 8,24 (s, 2H), 3,59 (s, 6H), 3,51 (s, 6H) 1 H NMR (300 MHz, CDCl 3 ) δ 8.24 (s, 2H), 3.59 (s, 6H), 3.51 (s, 6H)

(iii) Boc- NH- CH2-( 5- cvano)- 2- pvrimidin (iii) Boc-NH-CH2-(5-cvano)-2-pyrimidine

En blanding av f-butyl 2-amino-2-iminoetylkarbamat»AcOH (5,0 g, 23,8 mmol; se trinn (i) ovenfor) og l,3-bis(dimetylamino)-2-cyanotrimethinium perklorat (6,0 g, 23,8 mmol; se trinn (ii) ovenfor) i pyridin (300 ml) ble omrørt under nitrogen ved 70-75 °C i 16 timer og deretter oppvarmet under tilbakeløp i 6 timer. Blandingen ble avkjølt til romtemperatur og løsningsmidlet ble fjernet i vakuum. Residuet ble ekstrahert med en varm blanding (1:1) av etylacetat og kloroform, filtrert gjennom en liten pute av silika og konsentrert, hvilket gir råproduktet. Flash kromatografi på silika under eluering med kloroform ga 4,0 g (71%) av tittelforbindelsen som fargeløs olje, som stivnet ved henstand. A mixture of f-butyl 2-amino-2-iminoethylcarbamate»AcOH (5.0 g, 23.8 mmol; see step (i) above) and 1,3-bis(dimethylamino)-2-cyanotrimethinium perchlorate (6, 0 g, 23.8 mmol; see step (ii) above) in pyridine (300 mL) was stirred under nitrogen at 70-75 °C for 16 h and then heated under reflux for 6 h. The mixture was cooled to room temperature and the solvent was removed in vacuo. The residue was extracted with a hot mixture (1:1) of ethyl acetate and chloroform, filtered through a small pad of silica and concentrated to give the crude product. Flash chromatography on silica eluting with chloroform gave 4.0 g (71%) of the title compound as a colorless oil, which solidified on standing.

Sm.p.: 86-87 °C Melting point: 86-87 °C

Rf = 0,77 (silika, 3:2 Etylacetat/Kloroform) Rf = 0.77 (silica, 3:2 ethyl acetate/chloroform)

'H NMR (300 MHz, DMSO-</tf) 8 9,25 (s, 2H), 7,39 (bt, 1H), 4,39 (d, / = 6 Hz, 2H), 1,38 (s, 9H). 1 H NMR (300 MHz, DMSO-</tf) δ 9.25 (s, 2H), 7.39 (bt, 1H), 4.39 (d, / = 6 Hz, 2H), 1.38 ( pp, 9H).

<13>C NMR (750 MHz, DMSO-4$) 8 170,4,160,3,155,8, 115,2,106,9, 80,0,46,3, 28,1 APCI-MS: (M + 1) = 235 m/z <13>C NMR (750 MHz, DMSO-4$) δ 170.4,160.3,155.8, 115.2,106.9, 80.0.46.3, 28.1 APCI-MS: (M + 1) = 235 m/z

(iv) Boc- Aze- NH- CH2-(( 5- cvano)- 2- pvrimidinyl) (iv) Boc-Aze-NH-CH2-((5-cvano)-2-pvrimidinyl)

Boc-NH-CH2-(5-cyano)-2-pyrimidin (1,14 g, 4,87 mmol; se trinn (iii) ovenfor) ble oppløst i 50 ml EtOAc mettet med HCl(g) og fikk reagere i 1 time og konsentrert. Residuet ble oppløst i 20 ml DMF og avkjølt i et isbad. Diisopropyletylamin (3,5 ml, 0,020 mol), Boc-Aze-OH (1,08 g, 5,37 mmol) og HATU (2,80 g, 5,38 mmol) ble tilsatt og reaksjonsblandingen ble omrørt ved romtemperatur natten over. Løsningsmidlet ble avdampet og produktet ble renset ved preparativ RPLC ved anvendelse av CH3CN:0,1M NHiOAc (40:60). Acetonitrilet ble inndampet og det vandige laget ble ekstrahert tre ganger med EtOAc. De samlede organiske lag ble tørket (MgSO,*) og inndampet. Utbytte: 1,12 g (72%). Boc-NH-CH2-(5-cyano)-2-pyrimidine (1.14 g, 4.87 mmol; see step (iii) above) was dissolved in 50 mL EtOAc saturated with HCl(g) and allowed to react for 1 hour and concentrated. The residue was dissolved in 20 ml of DMF and cooled in an ice bath. Diisopropylethylamine (3.5 mL, 0.020 mol), Boc-Aze-OH (1.08 g, 5.37 mmol) and HATU (2.80 g, 5.38 mmol) were added and the reaction mixture was stirred at room temperature overnight . The solvent was evaporated and the product was purified by preparative RPLC using CH3CN:0.1M NHiOAc (40:60). The acetonitrile was evaporated and the aqueous layer was extracted three times with EtOAc. The combined organic layers were dried (MgSO 4 ) and evaporated. Yield: 1.12 g (72%).

<*>H NMR (400 MHz, CDCI3) 8 8,95 (s, 2H), 4,82 (d, 2H), 4,74 (m, 1H), 3,95 (m, 1H), 3,84 (m, 1H), 2,6-2,4 (m, 2H), 1,47 (s, 9H) <*>H NMR (400 MHz, CDCl 3 ) δ 8.95 (s, 2H), 4.82 (d, 2H), 4.74 (m, 1H), 3.95 (m, 1H), 3, 84 (m, 1H), 2.6-2.4 (m, 2H), 1.47 (s, 9H)

(v) Boc- Aze- NH- CH2-(( 5- amidino)- 2- pvrimidinvl) x HOAc (v) Boc-Aze-NH-CH2-((5-amidino)-2-pyrimidinyl) x HOAc

En løsning av Boc-Aze-NH-CH2-((5-cyano)-2-pyrimidinyl) (0,83 g, 2,6 mmol; se trinn (iv) ovenfor), N-acetylcystein (0,43 g, 2,6 mmol) og ammoniumacetat (0,60 g, 7,8 mmol) i 10 ml metanol ble oppvarmet ved 60°C under nitrogen i 2 dager. Løsningsmidlet ble avdampet og rå materiale ble renset ved preparativ RPLC ved anvendelse av en gradient av CH3CN:0,1M NH4OAC (5:95 til 100:0). Fraksjonene av interesse ble fryse tørket, hvilket gir 1,0 g (93%) av det ønskede materiale. A solution of Boc-Aze-NH-CH2-((5-cyano)-2-pyrimidinyl) (0.83 g, 2.6 mmol; see step (iv) above), N-acetylcysteine (0.43 g, 2.6 mmol) and ammonium acetate (0.60 g, 7.8 mmol) in 10 mL methanol was heated at 60 °C under nitrogen for 2 days. The solvent was evaporated and crude material was purified by preparative RPLC using a gradient of CH3CN:0.1M NH4OAC (5:95 to 100:0). The fractions of interest were freeze dried, yielding 1.0 g (93%) of the desired material.

<!>H NMR (300 MHz, D20, signaler utydelige grunnet HDO signal) 8 9.17 (s, 2H), 4,1-3,9 (m, 2H), 2,60 (m, 1H), 2,29 (m, 1H), 1,93 (s, 3H), 1,44 (s, 9H) (vi) Boc- Aze- NH- CH7- r( 5-( amino( trimetvlsilvletvlimino) metvl))- 2- pvrimidinvll Til en suspensjon av Boc-Aze-NH-CH2-((5-amidino)-2-pyrimidinyl) x HOAc (0,95 g, 2,41 mmol; se trinn (v) ovenfor) i 50 ml THF ble det tilsatt en løsning av Teoc-p-nitrofenylkarbonat (0,85 g, 3,0 mmol) og kaliumkarbonat (1,0 g, 7,2 mmol) i 10 ml vann. Blandingen ble omrørt i 24 timer, konsentrert og fordelt mellom vann og metylenklorid. Det organiske laget ble vasket to ganger med mettet vandig natriumbikarbonat, tørket (Na2S04) og inndampet. Råproduktet ble flash kromatografert på silikagel med heptan:EtOAc(l:l). Utbytte: 1,04 g (90%). <!>H NMR (300 MHz, D20, signals indistinct due to HDO signal) 8 9.17 (s, 2H), 4.1-3.9 (m, 2H), 2.60 (m, 1H), 2.29 (m, 1H), 1.93 (s, 3H), 1.44 (s, 9H) (vi) Boc-Aze-NH- CH7- r( 5-(amino(trimethylsilvletvlimino)metvl))- 2- pyrimidinevll To a suspension of Boc-Aze-NH-CH2-((5-amidino)-2-pyrimidinyl) x HOAc (0.95 g, 2.41 mmol; see step (v) above) in 50 mL of THF was added a solution of Teoc-p-nitrophenyl carbonate (0.85 g, 3.0 mmol) and potassium carbonate (1.0 g, 7.2 mmol) in 10 mL of water. The mixture was stirred for 24 hours, concentrated and partitioned between water and methylene chloride. The organic layer was washed twice with saturated aqueous sodium bicarbonate, dried (Na 2 SO 4 ) and evaporated. The crude product was flash chromatographed on silica gel with heptane:EtOAc (1:1). Yield: 1.04 g (90%).

<*>H NMR (300 MHz, CDCI3) 8 9,16 (s, 2H), 4,80 (d, 2H), 4,73 (m, 1H), 4,26 (m, 2H), 4,0-3,8 (m, 2H), 2,6-2,4 (m, 2H), 1,47 (s, 9H), 1,12 (m, 2H), 0,07 (s, 9H) <*>H NMR (300 MHz, CDCl 3 ) δ 9.16 (s, 2H), 4.80 (d, 2H), 4.73 (m, 1H), 4.26 (m, 2H), 4, 0-3.8 (m, 2H), 2.6-2.4 (m, 2H), 1.47 (s, 9H), 1.12 (m, 2H), 0.07 (s, 9H)

(vii) Ph( 3- Cl)( 5- OCHFz)-(/ g) CH( OH) C( Q)- Aze- NH- CH7- r( 5-( amino( trimetvlsilvletvlimino) metvl))- 2- pvrimidinvll (vii) Ph( 3- Cl)( 5- OCHFz)-(/ g) CH( OH) C( Q)- Aze- NH- CH7- r( 5-( amino(trimethylsilvletvlimino) metvl))- 2- pyrimidinevl

Boc-Aze->M-CH2-[(5-(ammo(trimetylsilyletylimino)metyl))-2-pyrimidin^ (0,209 g, 0,437 mmol; se trinn (vi) ovenfor) ble oppløst i 25 ml EtOAc mettet med HCl(g) og fikk reagere i 15 min. Løsningsmidlet ble avdampet og resten ble oppløst i 4 ml DMF. Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)OH (0,100 g, 0,396 mmol; se Eksempel l(viii) ovenfor), PyBOP (0,231 g, 0,444 mmol) og diisopropyletylamin (0,208 g, 1,61 mmol) ble tilsatt og blandingen ble omrørt i 80 min. Reaksjonsblandingen ble hellet i 100 ml vann og ekstrahert tre ganger med EtOAc. De samlede organiske lag ble vasket med saltvann, tørket (Na2S04) og inndampet. Råproduktet ble renset ved preparativ RPLC ved anvendelse av CH3CN:0,1M NH4OAC (1:1). Utbytte: 63 mg (26%). Boc-Aze->M-CH2-[(5-(ammo(trimethylsilylethylimino)methyl))-2-pyrimidine^ (0.209 g, 0.437 mmol; see step (vi) above) was dissolved in 25 mL of EtOAc saturated with HCl ( g) and allowed to react for 15 min. The solvent was evaporated and the residue was dissolved in 4 ml of DMF. Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)OH (0.100 g, 0.396 mmol; see Example 1(viii) above), PyBOP (0.231 g, 0.444 mmol) and diisopropylethylamine (0.208 g, 1.61 mmol) were added and the mixture was stirred for 80 min. The reaction mixture was poured into 100 mL of water and extracted three times with EtOAc. The combined organic layers were washed with brine, dried (Na 2 SO 4 ) and evaporated. The crude product was purified by preparative RPLC using CH3CN:0.1M NH4OAC (1:1). Yield: 63 mg (26%).

'H NMR (400 MHz, CDCI3, blanding av rotamerer) 6 9,3 (bred, 1H), 9,03 (s, 2H, mindre rotamer), 9,00 (s, 2H, hoved rotamer), 8,25 (m, 1H, hoved rotamer), 7,9 (bred, 1H), 7,80 (m, 1H, mindre rotamer), 7,2-6,9 (m, 3H), 6,50 (t, 1H), 5,14 (s, 1H, mindre rotamer), 5,08 (m, 1H, mindre rotamer), 4,94 (s, 1H, hoved rotamer), 4,80 (m, 1H, hoved rotamer), 4,7-4,4 (m, 2H), 4,3-3,9 (m, 3H), 3,74 (m, 1H, hoved rotamer), 2,7-2,1 (m, 2H), 1,03 (m, 2H), 0,01 (s, 9H) 1H NMR (400 MHz, CDCl3, mixture of rotamers) δ 9.3 (broad, 1H), 9.03 (s, 2H, minor rotamer), 9.00 (s, 2H, major rotamer), 8.25 (m, 1H, major rotamer), 7.9 (broad, 1H), 7.80 (m, 1H, minor rotamer), 7.2-6.9 (m, 3H), 6.50 (t, 1H ), 5.14 (s, 1H, minor rotamer), 5.08 (m, 1H, minor rotamer), 4.94 (s, 1H, major rotamer), 4.80 (m, 1H, major rotamer), 4.7-4.4 (m, 2H), 4.3-3.9 (m, 3H), 3.74 (m, 1H, main rotamer), 2.7-2.1 (m, 2H) , 1.03 (m, 2H), 0.01 (s, 9H)

(viii) Ph( 3- Cl)( 5- OCHF7)- r/ ?) CH( OH) C( 0)- Aze- NH- CH2-(( 5- amidino)- 2- pvrimidinvl) x (viii) Ph( 3- Cl)( 5- OCHF7)- r/ ?) CH( OH) C( 0)- Aze- NH- CH2-(( 5- amidino)- 2- pvrimidinvl) x

TFA TFA

Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)-Aze-NH-CH2-[(5-(amino(trimetyl-silyletylimino)metyl))-2-pyrimidinyl] (21 mg, 0,034 mmol; se trinn (vii) ovenfor) ble oppløst i 0,5 ml metylenklorid og avkjølt i et isbad. TFA (2 ml) ble tilsatt og blandingen ble omrørt i 60 min og deretter konsentrert. Produktet ble frysetørket fra vann og acetonitril. Utbytte: 20 mg (100%). Renhet: 100%. Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)-Aze-NH-CH2-[(5-(amino(trimethyl-silylethylimino)methyl))-2-pyrimidinyl] (21 mg, 0.034 mmol; see step (vii) above) was dissolved in 0.5 mL of methylene chloride and cooled in an ice bath. TFA (2 mL) was added and the mixture was stirred for 60 min and then concentrated. The product was lyophilized from water and acetonitrile Yield: 20 mg (100%) Purity: 100%

'H NMR (400 MHz, CD3OD, blanding av rotamer signaler obscured by HDO signal) 8 9,08 (s, 2H), 7,4-7,1 (m, 3H), 6,88 (t, 1H, hoved rotamer), 6,85 (t, 1H, mindre rotamer), 5,30 (m, 1H, mindre rotamer), 5,22 (s, 1H, mindre rotamer), 5,20 (s, 1H, hoved rotamer), 4,73 (m, 1H, hoved rotamer), 4,34 (m, 1H, rotamer), 4,21 (m, 1H, rotamer), 4,15-3,95 (m, 2H, rotamerer), 2,73 (m, 1H, rotamer), 2,57 (m, 1H, rotamer), 2,45-2,25 (m, 2H, rotamerer) 'H NMR (400 MHz, CD3OD, mixture of rotamer signals obscured by HDO signal) 8 9.08 (s, 2H), 7.4-7.1 (m, 3H), 6.88 (t, 1H, main rotamer), 6.85 (t, 1H, minor rotamer), 5.30 (m, 1H, minor rotamer), 5.22 (s, 1H, minor rotamer), 5.20 (s, 1H, major rotamer) , 4.73 (m, 1H, main rotamer), 4.34 (m, 1H, rotamer), 4.21 (m, 1H, rotamer), 4.15-3.95 (m, 2H, rotamers), 2.73 (m, 1H, rotamer), 2.57 (m, 1H, rotamer), 2.45-2.25 (m, 2H, rotamer)

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 173,0,172,6,172,1,171,0,163,4. <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 173.0.172.6.172.1.171.0.163.4.

APCI-MS: (M + 1) = 469/471 m/z APCI-MS: (M + 1) = 469/471 m/z

Eksempel 37 Example 37

Ph( 3- Cl¥5- OCHF?)-(/ g) CH( OmC( 0)- Aze- NH- CH7-(( 5- metoksvamidino)- 2- pvrimidinvn Ph( 3- Cl¥5- OCHF?)-(/ g) CH( OmC( 0)- Aze- NH- CH7-(( 5- methoxyvamidino)- 2- pvrimidinvn

(i) Ph( 3- Cn( 5- OCHF7)- r/ ?JCH( OH) C( 0)- Aze- NH- CH?- r( 5-( metoksvamino-( trimetvlsilvletylimino) metyl))- 2- pvrimidinvl1 (i) Ph( 3- Cn( 5- OCHF7)- r/ ?JCH( OH) C( 0)- Aze- NH- CH?- r( 5-( methoxyamino-(trimethylsilvethylimino) methyl))- 2- pyrimidinev1

En suspensjon av Ph(3-Cl)(5-OCHF2H/?)CH(OH)C(0)-Aze-NH-CH2-[(5-(amino(trimetylsilyletylimino)metyl))-2-pyrimidinyl] (40 mg, 0,065 mmol; se Eksempel 36(vii) ovenfor) og O-metylhydroksylamin (33 mg, 0,40 mmol) i 3 ml acetonitril ble oppvarmet ved 70°C i 3 timer. Blandingen ble fordelt mellom vann og EtOAc. Det vandige laget ble ekstrahert to ganger med EtOAc og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Utbytte: 33 mg (79%). A suspension of Ph(3-Cl)(5-OCHF2H/?)CH(OH)C(0)-Aze-NH-CH2-[(5-(amino(trimethylsilylethylimino)methyl))-2-pyrimidinyl] (40 mg, 0.065 mmol; see Example 36(vii) above) and O-methylhydroxylamine (33 mg, 0.40 mmol) in 3 mL of acetonitrile was heated at 70°C for 3 hours. The mixture was partitioned between water and EtOAc. The aqueous layer was extracted twice with EtOAc and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 33 mg (79%).

<*>H NMR (400 MHz, CDC13, blanding av rotamerer) 88,76 (s 2H, hoved rotamer), 8,70 (s, 2H, rotamer), 8,18 (m, 1H), 7,62 (s, 1H), 7,4-6,9 (m, 4H), 6,50 (bt, 1H), 5,3-4,5 (m, 4H), 4,2-4,05 (m, 3H), 3,96 (s, 3H), 3,68 (m, 1H), 2,8-2,2 (m, 2H), 2,1 (bred, 1H), 0,96 (m, 2H), 0,01 (s, 9H) <*>H NMR (400 MHz, CDCl 3 , mixture of rotamers) 88.76 (s 2H, main rotamer), 8.70 (s, 2H, rotamer), 8.18 (m, 1H), 7.62 ( s, 1H), 7.4-6.9 (m, 4H), 6.50 (bt, 1H), 5.3-4.5 (m, 4H), 4.2-4.05 (m, 3H), 3.96 (s, 3H), 3.68 (m, 1H), 2.8-2.2 (m, 2H), 2.1 (broad, 1H), 0.96 (m, 2H ), 0.01 (s, 9H)

(ii) Ph( 3- Cl)( 5- OCHF?)- r/ g) CH( OH) C( 0)- Aze- NH- CHz-(( 5- metoksv-amidino)- 2- pvrimidinyl) (ii) Ph( 3- Cl)( 5- OCHF?)- r/ g) CH( OH) C( 0)- Aze- NH- CHz-(( 5- methoxysv-amidino)- 2- pvrimidinyl)

Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-NH-CH2-[(5-(metoksyamino-(trimetylsilyletylimino)metyl))-2-pyrimidinyl] (33 mg, 0,052 mmol; se trinn (i) ovenfor) ble oppløst i 0,5 ml metylenklorid og avkjølt i et isbad. TFA (2 ml) ble tilsatt og blandingen ble omrørt i 2 timer og deretter konsentrert. Produktet ble fryse tørket fra vann og acetonitril. Utbytte: 31 mg (81%). Renhet: 100%. Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-NH-CH2-[(5-(Methoxyamino-(trimethylsilylethylimino)methyl))-2-pyrimidinyl] (33 mg, 0.052 mmol; see step (i) above) was dissolved in 0.5 mL of methylene chloride and cooled in an ice bath. TFA (2 mL) was added and the mixture was stirred for 2 h and then concentrated. The product was freeze dried from water and acetonitrile. Yield: 31 mg (81%). Purity: 100%.

<*>H NMR (400 MHz, CD3OD, blanding av rotamer signaler obscured by HDO signal) 88,96 (s, 2H, rotamer), 8,94 (s, 2H, rotamer), 7,4-7,3 (m, 1H), 7,2-7,1 (m, 2H), 6,88 (t, 1H, rotamer), 6,85 (t, 1H, rotamer), 5,29 (m, 1H, rotamer), 5,24 (s, 1H, rotamer), 5,20 (s, 1H, rotamer), 4,75-4,55 (m, 2H), 4,33 (m, 1H, rotamer), 4,19 (m, 1H, rotamer), 4,15-3,95 (m, 2H, rotamerer), 3,88 (s, 3H, rotamer), 3,86 (s, 3H, rotamer), 2,72 (m, 1H, rotamer), 2,56 (m, 1H, rotamer), 2,45-2,25 (m, 2H, rotamerer) <*>H NMR (400 MHz, CD3OD, mixture of rotamer signals obscured by HDO signal) 88.96 (s, 2H, rotamer), 8.94 (s, 2H, rotamer), 7.4-7.3 ( m, 1H), 7.2-7.1 (m, 2H), 6.88 (t, 1H, rotamer), 6.85 (t, 1H, rotamer), 5.29 (m, 1H, rotamer) , 5.24 (s, 1H, rotamer), 5.20 (s, 1H, rotamer), 4.75-4.55 (m, 2H), 4.33 (m, 1H, rotamer), 4.19 (m, 1H, rotamer), 4.15-3.95 (m, 2H, rotamer), 3.88 (s, 3H, rotamer), 3.86 (s, 3H, rotamer), 2.72 (m , 1H, rotamer), 2.56 (m, 1H, rotamer), 2.45-2.25 (m, 2H, rotamer)

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 172,8,172,6,172,1,171,8,167,8,167,7,155,1,152,3,152,1 <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 172,8,172,6,172,1,171,8,167,8,167,7,155,1,152,3,152,1

APCI-MS: (M + 1) = 499/501 m/z APCI-MS: (M + 1) = 499/501 m/z

Eksempel 38 Example 38

Ph( 3- Cl)( 5- OCHFz)-(/ g) CH( OH) C( Q)- Aze- Pab( 3- F) Ph( 3- Cl)( 5- OCHFz)-(/ g) CH( OH) C( Q)- Aze- Pab( 3- F)

(i) 2- fluor- 4- vinylbenzonitril (i) 2-fluoro-4-vinylbenzonitrile

En løsning av 4-brom-2-fluorbenzonitril (4,92 g, 0,0246 mol), vinyltributyltin (0,78 g, 0,246 mol) og tetrakistrifenylfosfin (0,67 g, 0,58 mmol) i 250 ml toluen ble tilbakeløpskokt under nitrogen natten over. Løsningsmidlet ble avdampet og residuet ble flash kromatografert på silikagel med heptamC^Ch (1:1) til ren CH2CI2. En fargeløs olje ble oppnådd som krystalliserte. Utbytte: 3,0 g (82%). A solution of 4-bromo-2-fluorobenzonitrile (4.92 g, 0.0246 mol), vinyltributyltin (0.78 g, 0.246 mol) and tetrakistriphenylphosphine (0.67 g, 0.58 mmol) in 250 mL of toluene was refluxed under nitrogen overnight. The solvent was evaporated and the residue was flash chromatographed on silica gel with heptamCl 2 Cl 2 (1:1) to pure CH 2 Cl 2 . A colorless oil was obtained which crystallized. Yield: 3.0 g (82%).

'H NMR (300 MHz, CDC13) 8 7,56 (m, 1H), 7,3-7,2 (m, 2H), 6,69 (m, 1H), 5,89 (d, 1H), 5,51 (d, 1H) 1 H NMR (300 MHz, CDCl 3 ) δ 7.56 (m, 1H), 7.3-7.2 (m, 2H), 6.69 (m, 1H), 5.89 (d, 1H), 5.51 (d, 1H)

(ii) 2- fluor- 4- hvdroksvmetylbenzonitril (ii) 2-fluoro-4-hydroxymethylbenzonitrile

Til en avkjølt løsning (-78°C) av 2-fluor-4-vinylbenzonitril (1,3 g, 8,8 mmol; se trinn (i) ovenfor) i 40 ml CH2CI2 og 5 ml metanol ble det boblet ozon (50 L/h, 29 g/m<3>) i 30 min. Argon ble deretter boblet gjennom for å fjerne overskudd av ozon. Natrium-borhydrid (0,67 g, 0,018 mol) ble tilsatt og kjølebadet ble fjernet. Blandingen ble omrørt og fikk reagere i 1 time. Blandingen ble inndampet og 2M HC1 ble tilsatt. Blandingen ble ekstrahert to ganger med dietyleter og den samlede eterfraksjon ble tørket (Na2SC«4) og inndampet. Råproduktet krystallisert. Utbytte: 1,1 g (81%). To a cooled solution (-78°C) of 2-fluoro-4-vinylbenzonitrile (1.3 g, 8.8 mmol; see step (i) above) in 40 mL CH 2 Cl 2 and 5 mL methanol was bubbled ozone (50 L/h, 29 g/m<3>) for 30 min. Argon was then bubbled through to remove excess ozone. Sodium borohydride (0.67 g, 0.018 mol) was added and the cooling bath was removed. The mixture was stirred and allowed to react for 1 hour. The mixture was evaporated and 2M HCl was added. The mixture was extracted twice with diethyl ether and the combined ether fraction was dried (Na 2 SC 4 ) and evaporated. The crude product crystallized. Yield: 1.1 g (81%).

<*>H NMR (300 MHz, CDC13) 8 7,59 (m, 1H), 7,3-7,2 (m, 2H), 4,79 (d, 2H), 2,26 (t, 1H) <*>H NMR (300 MHz, CDCl 3 ) δ 7.59 (m, 1H), 7.3-7.2 (m, 2H), 4.79 (d, 2H), 2.26 (t, 1H )

(iii) 4- cvano- 3- fluorbenzyl metansulfonat (iii) 4-cvano-3-fluorobenzyl methanesulfonate

2-fluor-4-hydroksymetylbenzonitril (1,3 g, 8,6 mmol; se trinn (ii) ovenfor) ble oppløst i 50 ml CH2CI2 og avkjølt på et isbad. Trietylamin (0,87 g, 8,6 mmol) og metansulfonylklorid (0,99 g, 8,7 mmol) ble tilsatt. Etter omrøring i 1,5 timer ble reaksjonsblandingen vasket med IM HC1. Den organiske fasen ble tørket (Na2SC<4) og inndampet. Produktet kunne anvendes uten rensning. Utbytte av en fargeløs olje: 1,8 g (92%). 2-Fluoro-4-hydroxymethylbenzonitrile (1.3 g, 8.6 mmol; see step (ii) above) was dissolved in 50 mL CH 2 Cl 2 and cooled in an ice bath. Triethylamine (0.87 g, 8.6 mmol) and methanesulfonyl chloride (0.99 g, 8.7 mmol) were added. After stirring for 1.5 hours, the reaction mixture was washed with 1M HCl. The organic phase was dried (Na2SO4) and evaporated. The product could be used without cleaning. Yield of a colorless oil: 1.8 g (92%).

'H NMR (400 MHz, CDCI3) 8 7,66 (m, 1H), 7,35-7,3 (m, 2H), 5,26 (s, 2H), 3,07 (s, 3H) 1 H NMR (400 MHz, CDCl 3 ) δ 7.66 (m, 1H), 7.35-7.3 (m, 2H), 5.26 (s, 2H), 3.07 (s, 3H)

(iv) 4- Azidometvl- 2- lfuorbenzonitril (iv) 4-Azidomethyl-2-fluorobenzonitrile

Til en is avkjølt løsning av 4-cyano-3-fluorbenzylmetansulfonat (1,8 g, 7,9 mmol; se trinn (iii) ovenfor) ble det tilsatt natriumazid (0,80 g, 0,012 mol). Blandingen ble omrørt natten over og deretter hellet i 200 ml vann og ekstrahert tre ganger med dietyleter. Den samlede eteriske fasen ble vasket fem ganger med vann, tørket (Na2S04) og inndampet. Rå fargeløs olje kunne anvendes uten ytterligere rensning. Utbytte: 1,2 g (87%). To an ice-cooled solution of 4-cyano-3-fluorobenzyl methanesulfonate (1.8 g, 7.9 mmol; see step (iii) above) was added sodium azide (0.80 g, 0.012 mol). The mixture was stirred overnight and then poured into 200 ml of water and extracted three times with diethyl ether. The combined ether phase was washed five times with water, dried (Na 2 SO 4 ) and evaporated. Crude colorless oil could be used without further purification. Yield: 1.2 g (87%).

<!>H NMR (300 MHz, CDCI3) 8 7,64 (m, 1H), 7,25-7,18 (m, 2H), 4,47 (s, 2H) <!>H NMR (300 MHz, CDCl 3 ) δ 7.64 (m, 1H), 7.25-7.18 (m, 2H), 4.47 (s, 2H)

(v) 4- aminometvl- 2- lfuorbenzonitril (v) 4-aminomethyl-2-fluorobenzonitrile

Til en suspensjon av tinnkloriddihydrat (0,45 g, 2,4 mmol) i 20 ml acetonitril under omrøring ble det tilsatt tiofenol (1,07 g, 9,7 mmol) og trietylamin (0,726 g, 7,17 mmol). Deretter ble det tilsatt en løsning av 4-azidometyl-2-fluorbenzonitril (0,279 g, 1,58 mmol; se trinn (iv) ovenfor) i noen få mL av acetonitril. Etter 1,5 timer, ble azidet oppbrukt og løsningsmidlet ble avdampet. Residuet ble oppløst i metylenklorid og vasket tre ganger med 2M NaOH. Den organiske fasen ble ekstrahert to ganger med IM HC1. Den samlede sure vandige fasen ble vasket med metylenklorid og deretter gjort alkalisk med 2M NaOH og ekstrahert tre ganger med metylenklorid. Den organiske fasen ble tørket (Na2S04) og inndampet, hvilket gir 0,172 g (72%) av den ønskede sub-tittelforbindelsen som kunne anvendes uten rensning. To a suspension of stannous chloride dihydrate (0.45 g, 2.4 mmol) in 20 mL of acetonitrile with stirring was added thiophenol (1.07 g, 9.7 mmol) and triethylamine (0.726 g, 7.17 mmol). Then a solution of 4-azidomethyl-2-fluorobenzonitrile (0.279 g, 1.58 mmol; see step (iv) above) in a few mL of acetonitrile was added. After 1.5 hours, the azide was consumed and the solvent was evaporated. The residue was dissolved in methylene chloride and washed three times with 2M NaOH. The organic phase was extracted twice with IM HCl. The combined acidic aqueous phase was washed with methylene chloride and then made alkaline with 2M NaOH and extracted three times with methylene chloride. The organic phase was dried (Na 2 SO 4 ) and evaporated to give 0.172 g (72%) of the desired sub-title compound which could be used without purification.

'H NMR (400 MHz, CDC13) 8 7,58 (m, 1H), 7,3-7,2 (m, 2H), 3,98 (s, 2H), 1,55-1,35 (bred, 2H) 1 H NMR (400 MHz, CDCl 3 ) δ 7.58 (m, 1H), 7.3-7.2 (m, 2H), 3.98 (s, 2H), 1.55-1.35 (broad , 2H)

(vi) Boc- Aze- NHCH7- Ph( 3- F. 4- CN) (vi) Boc- Aze- NHCH7- Ph( 3- F. 4- CN)

Til en is avkjølt løsning av Boc-Aze-OH (0,194 g, 0,96 mmol) i 5 ml DMF ble det tilsatt TBTU (0,50 g, 9,6 mmol). Etter 30 min ble en ytterligere løsning, omfattende 4-aminometyl-2-fluorbenzonitril (0,17 g, 0,81 mmol; se trinn (v) ovenfor) og diiisopropyletylamin (0,326 g, 2,53 mmol) i 7 ml DMF tilsatt. Den resulterende løsning ble omrørt natten over ved romtemperatur. Løsningsmidlet ble avdampet og produktet ble renset ved preparativ RPLC ved anvendelse av CH3CN:0,1M NH4OAC (50:50). Frysetørking ga 0,237 g (74%) av den ønskede sub-tittelforbindelsen. To an ice-cooled solution of Boc-Aze-OH (0.194 g, 0.96 mmol) in 5 mL DMF was added TBTU (0.50 g, 9.6 mmol). After 30 min, an additional solution comprising 4-aminomethyl-2-fluorobenzonitrile (0.17 g, 0.81 mmol; see step (v) above) and diisopropylethylamine (0.326 g, 2.53 mmol) in 7 mL of DMF was added . The resulting solution was stirred overnight at room temperature. The solvent was evaporated and the product was purified by preparative RPLC using CH3CN:0.1M NH4OAC (50:50). Freeze drying gave 0.237 g (74%) of the desired sub-title compound.

'H NMR (300 MHz, CD3OD) 8 7,70 (m, 1H), 7,35-7,25 (m, 2H), 4,65-4,35 (m, 3H), 4,0-3,85 (m, 2H), 2,51 (m, 1H), 2,19 (m, 1H), 1,40 (s, 9H) (vii) Ph( 3- Cl)( 5- OCHF7)- r/ g) CH( OH) C( 0)- Aze- NHCHz- Ph( 3- F. 4- CN) Boc-Aze-NHCH2-Ph(3-F, 4-CN) (0,118 g, 0,354 mmol; fra trinn (vi) ovenfor) ble oppløst i 30 ml EtOAc mettet med HCl(g). Reaksjonsblandingen ble omrørt i 20 min og inndampet. Det resulterende dihydroklorid og HATU (0,152 g, 0,400 mmol) ble oppløst i 5 ml DMF. Løsningen ble satt til en is avkjølt løsning av Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)OH (0,101 g, 0,400 mmol; se Eksempel l(viii) ovenfor) i 5 ml DMF. Reaksjonsblandingen ble omrørt natten over ved omgivelsestemperatur. Løsningsmidlet ble avdampet og produktet ble renset ved preparativ RPLC med CH3CN:0,1M NH4OAC (50:50). Frysetørking ga 0,130 g (77%) av den ønskede sub-tittelforbindelsen. 1 H NMR (300 MHz, CD 3 OD) δ 7.70 (m, 1H), 7.35-7.25 (m, 2H), 4.65-4.35 (m, 3H), 4.0-3 .85 (m, 2H), 2.51 (m, 1H), 2.19 (m, 1H), 1.40 (s, 9H) (vii) Ph( 3- Cl)( 5- OCHF7)- r / g) CH( OH) C( 0)- Aze- NHCHz- Ph( 3- F. 4- CN) Boc-Aze-NHCH2-Ph(3-F, 4-CN) (0.118 g, 0.354 mmol; from step (vi) above) was dissolved in 30 mL of EtOAc saturated with HCl(g). The reaction mixture was stirred for 20 min and evaporated. The resulting dihydrochloride and HATU (0.152 g, 0.400 mmol) were dissolved in 5 mL of DMF. The solution was added to an ice-cooled solution of Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)OH (0.101 g, 0.400 mmol; see Example 1(viii) above) in 5 mL DMF. The reaction mixture was stirred overnight at ambient temperature. The solvent was evaporated and the product was purified by preparative RPLC with CH3CN:0.1M NH4OAC (50:50). Freeze-drying gave 0.130 g (77%) of the desired sub-title compound.

'H NMR (500 MHz, CD3OD blanding av rotamerer) 8 7,7-7,6 (m, 1H), 7,35-7,1 (m, 5H), 6,88 (t, 1H, rotamer), 6,86 (t, 1H, rotamer), 5,25-5,1 (m, 1H pluss mindre rotamer fra det 1H NMR (500 MHz, CD3OD mixture of rotamers) δ 7.7-7.6 (m, 1H), 7.35-7.1 (m, 5H), 6.88 (t, 1H, rotamer), 6.86 (t, 1H, rotamer), 5.25-5.1 (m, 1H plus minor rotamer from the

følgende proton), 4,80 (m, 1H, hoved rotamer), 4,6-4,4 (m, 2H), 4,36 (m, 1H, hoved rotamer), 4,18 (m, 1H, hoved rotamer), 4,07 (m, 1H, mindre rotamer), 3,98 (m, 1H, mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,53 (m, 1H, hoved rotamer), 2,29 (m, 1H, hoved rotamer), 2,16 (m, 1H, mindre rotamer) following proton), 4.80 (m, 1H, main rotamer), 4.6-4.4 (m, 2H), 4.36 (m, 1H, main rotamer), 4.18 (m, 1H, main rotamer), 4.07 (m, 1H, minor rotamer), 3.98 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.53 (m, 1H, major rotamer) , 2.29 (m, 1H, major rotamer), 2.16 (m, 1H, minor rotamer)

(viii) Ph( 3- Cl)( 5- OCHF7)- r/ g) CH( OH) C( Q)- Aze- Pab( 3- F) (viii) Ph( 3- Cl)( 5- OCHF7)- r/ g) CH( OH) C( Q)- Aze- Pab( 3- F)

Ph(3-Cl)(5-OCHF2)-r/?>)CH(OH)C(0)-Aze-NHCH2-Ph(3-F, 4-CN) (0,130 g, 0,278 mmol; se trinn (vii) ovenfor) ble oppløst i 80 ml etanol mettet med HCl(g). Blandingen fikk reagere ved romtemperatur natten over. Løsningsmidlet ble avdampet og residuet ble gjenoppløst i 100 ml etanol mettet med NH3(g). Reaksjonen fikk forløpe langsomt ved romtemperatur i to dager. Temperaturen ble hevet til 50°C og reaksjonen fortsatt i ytterligere 3 dager. Utgangsmaterialet ble oppbrukt og løsningsmidlet ble avdampet. Produktet ble renset ved preparativ RPLC og frysetørket, hvilket gir 17 mg (13%) av tittelforbindelsen som dens HOAc salt. Ph(3-Cl)(5-OCHF2)-r/?>)CH(OH)C(0)-Aze-NHCH2-Ph(3-F, 4-CN) (0.130 g, 0.278 mmol; see step ( vii) above) was dissolved in 80 ml of ethanol saturated with HCl(g). The mixture was allowed to react at room temperature overnight. The solvent was evaporated and the residue redissolved in 100 ml of ethanol saturated with NH3(g). The reaction was allowed to proceed slowly at room temperature for two days. The temperature was raised to 50°C and the reaction continued for a further 3 days. The starting material was used up and the solvent was evaporated. The product was purified by preparative RPLC and lyophilized to give 17 mg (13%) of the title compound as its HOAc salt.

<*>H NMR (600 MHz, CD3OD blanding av rotamerer) 8 7,65-7,6 (m, 1H), 7,4-7,3 (m, 3H), 7,25-7,1 (m, 2H), 7,15-6,7 (m, 1H), 5,25-5,1 (m, 1H pluss mindre rotamer av de følgende proton), 4,8 (m, 1H, hoved rotamer delvis hidden by CD3OH), 4,6-3,95 (m, 4H), 2,69 (m, 1H, mindre rotamer), 2,56 (m, 1H, hoved rotamer), 2,28 (m, 1H, hoved rotamer), 2,14 (m, 1H, mindre rotamer), 1,90 (s, 3H) <*>H NMR (600 MHz, CD3OD mixture of rotamers) δ 7.65-7.6 (m, 1H), 7.4-7.3 (m, 3H), 7.25-7.1 (m , 2H), 7.15-6.7 (m, 1H), 5.25-5.1 (m, 1H plus minor rotamer of the following protons), 4.8 (m, 1H, main rotamer partially hidden by CD3OH), 4.6-3.95 (m, 4H), 2.69 (m, 1H, minor rotamer), 2.56 (m, 1H, major rotamer), 2.28 (m, 1H, major rotamer ), 2.14 (m, 1H, minor rotamer), 1.90 (s, 3H)

<l3>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 180,6,173,4,173,1,172,9,164,5,162,3,159,8 <l3>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 180,6,173,4,173,1,172,9,164,5,162,3,159,8

APCI-MS: (M + 1) = 485/487 m/z APCI-MS: (M + 1) = 485/487 m/z

Eksempel 39 Example 39

Ph( 3- Cl)( 5- OCHF7)-(/ ?) CH( OH) C( Q)- Aze- Pab( 2. 6- diF) Ph( 3- Cl)( 5- OCHF7)-(/ ?) CH( OH) C( Q)- Aze- Pab( 2. 6- diF)

(i) 2. 6- difluor- 4r( metvlsulfinvl)( metvltio) metyllbenzonitril (i) 2. 6-difluoro-4r(methylsulfinyl)(methylthio)methylbenzonitrile

(Metylsulfinyl)(metyltio)metan (7,26 g, 0,0584 mol) ble oppløst i 100 ml tørr THF under argon og ble avkjølt til -78°C. Butyllitium i heksan (16 ml 1,6M, 0,0256 mol) ble tilsatt dråpevis med omrøring. Blandingen ble omrørt i 15 min. I mens ble, en løsning av 3,4,5-trifluorbenzonitril (4,0 g, 0,025 mmol) i 100 ml tørr THF avkjølt til -78°C under argon og førstnevnte løsning ble tilsatt gjennom en kanyle til den sistnevnte løsning over en periode på 35 min. Etter 30 min, ble kjølebadet fjernet og når reaksjonen hadde nådd romtemperatur ble den hellet i 400 ml vann. THF ble inndampet og det gjenværende vandige lag ble ekstrahert tre ganger med dietyleter. Den samlede eter fasen ble vasket med vann, tørket (Na2S04) og inndampet. Utbytte: 2,0 g (30%). (Methylsulfinyl)(methylthio)methane (7.26 g, 0.0584 mol) was dissolved in 100 mL of dry THF under argon and cooled to -78°C. Butyllithium in hexane (16 mL 1.6M, 0.0256 mol) was added dropwise with stirring. The mixture was stirred for 15 min. Meanwhile, a solution of 3,4,5-trifluorobenzonitrile (4.0 g, 0.025 mmol) in 100 mL dry THF was cooled to -78 °C under argon and the former solution was added through a cannula to the latter solution over a period of 35 min. After 30 min, the cooling bath was removed and when the reaction had reached room temperature it was poured into 400 ml of water. The THF was evaporated and the remaining aqueous layer was extracted three times with diethyl ether. The combined ether phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 2.0 g (30%).

<!>H NMR (500 MHz, CDC13) 8 7,4-7,25 (m, 2H), 5,01 (s, 1H, diasteromer), 4,91 (s, 1H, diasteromer), 2,88 (s, 3H, diasteromer), 2,52 (s, 3H, diasteromer), 2,49 (s, 3H, diasteromer), 2,34 (s, 3H, diasteromer), 1,72 (bred, 1H) <!>H NMR (500 MHz, CDCl 3 ) δ 7.4-7.25 (m, 2H), 5.01 (s, 1H, diastereomers), 4.91 (s, 1H, diastereomers), 2.88 (s, 3H, diastereomers), 2.52 (s, 3H, diastereomers), 2.49 (s, 3H, diastereomers), 2.34 (s, 3H, diastereomers), 1.72 (broad, 1H)

(ii) 2. 6- difluor- 4- formvlbenzonitril (ii) 2. 6-difluoro-4-formylbenzonitrile

2,6-difluor-4[(metylsulfinyl)(metyltio)metyl]benzonitril (2,17 g, 8,32 mmol; se trinn (i) ovenfor) ble oppløst i 90 ml THF og 3,5 ml konsentrert svovelsyre ble tilsatt. Blandingen fikk stå ved romtemperatur i 3 dager og deretter hellet i 450 ml vann. Ekstraksjon tre ganger med EtOAc og den samlede eteriske fase ble vasket to ganger med vandig natriumbikarbonat og med saltvann, tørket (Na2SC<4) og inndampet. Utbytte: 1,36 g (98%). Stillingen til formylgruppen ble etablert ved CNMR. Signalet fra fluorerte karbonatomer ved 162,7 ppm viste forventet koblingsmønster med to koblingskonstanter i 2,6-Difluoro-4[(methylsulfinyl)(methylthio)methyl]benzonitrile (2.17 g, 8.32 mmol; see step (i) above) was dissolved in 90 mL of THF and 3.5 mL of concentrated sulfuric acid was added . The mixture was allowed to stand at room temperature for 3 days and then poured into 450 ml of water. Extracted three times with EtOAc and the combined ethereal phase was washed twice with aqueous sodium bicarbonate and with brine, dried (Na2SO4) and evaporated. Yield: 1.36 g (98%). The post for the formyl group was established at the CNMR. The signal from fluorinated carbon atoms at 162.7 ppm showed the expected coupling pattern with two coupling constants in

størrelse 260 Hz og 6,3 Hz henholdsvis svarende til en ipso og en meta kobling fra fluoratomer. size 260 Hz and 6.3 Hz respectively corresponding to an ipso and a meta coupling from fluorine atoms.

<!>H NMR (400 MHz, CDC13) 8 10,35 (s, 1H), 7,33 (m, 2H) <!>H NMR (400 MHz, CDCl 3 ) δ 10.35 (s, 1H), 7.33 (m, 2H)

(iii) 2, 6- difluor- 4- hydroksymetylbenzonitril (iii) 2,6-difluoro-4-hydroxymethylbenzonitrile

2,6-difluor-4-formylbenzonitril (1,36 g, 8,13 mmol; se trinn (ii) ovenfor) ble oppløst i 25 ml metanol og avkjølt på et isbad. Natrium-borhydrid (0,307 g, 8,12 mmol) ble tilsatt i porsjoner med omrøring og reaksjonen ble latt stå i 65 min. Løsningsmidlet ble avdampet og residuet ble fordelt mellom dietyleter og vandig natriumbikarbonat. Det eteriske lag ble vasket med mer vandig natriumbikarbonat og saltvann, tørket (Na2S04) og inndampet. Råproduktet krystallisert fort og kunne anvendes uten ytterligere rensning. Utbytte: 1,24 g (90%). 2,6-Difluoro-4-formylbenzonitrile (1.36 g, 8.13 mmol; see step (ii) above) was dissolved in 25 mL of methanol and cooled in an ice bath. Sodium borohydride (0.307 g, 8.12 mmol) was added in portions with stirring and the reaction was allowed to stand for 65 min. The solvent was evaporated and the residue partitioned between diethyl ether and aqueous sodium bicarbonate. The ethereal layer was washed with more aqueous sodium bicarbonate and brine, dried (Na 2 SO 4 ) and evaporated. The crude product crystallized quickly and could be used without further purification. Yield: 1.24 g (90%).

'H NMR (400 MHz, CDC13) 8 7,24 (m, 2H), 4,81 (s, 2H), 2,10 (bred, 1H) 1 H NMR (400 MHz, CDCl 3 ) δ 7.24 (m, 2H), 4.81 (s, 2H), 2.10 (broad, 1H)

(iv) 4- cvano- 2. 6- difluorbenzyl metansulfonat (iv) 4-cvano-2.6-difluorobenzyl methanesulfonate

Til en is avkjølt løsning av 2,6-difluor-4-hydroksymetylbenzonitril (1,24 g, 7,32 mmol; se trinn (iii) ovenfor) og metansulfonylklorid (0,93 g, 8,1 mmol) i 60 ml metylenklorid ble det tilsatt trietylamin (0,81 g, 8,1 mmol) med omrøring. Etter 3 timer ved 0°C, ble blandingen vasket to ganger med IM HC1 og én gang med vann, tørket (Na2SC«4) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 1,61 g (89%). To an ice-cooled solution of 2,6-difluoro-4-hydroxymethylbenzonitrile (1.24 g, 7.32 mmol; see step (iii) above) and methanesulfonyl chloride (0.93 g, 8.1 mmol) in 60 mL of methylene chloride triethylamine (0.81 g, 8.1 mmol) was added with stirring. After 3 hours at 0°C, the mixture was washed twice with 1M HCl and once with water, dried (Na 2 SC 4 ) and evaporated. The product could be used without further purification. Yield: 1.61 g (89%).

'H NMR (300 MHz, CDC13) 8 7,29 (m, 2H), 5,33 (s, 2H), 3,07 (s, 3H) 1 H NMR (300 MHz, CDCl 3 ) δ 7.29 (m, 2H), 5.33 (s, 2H), 3.07 (s, 3H)

(v) 4- Azidometyl- 2. 6- difluorbenzonitril (v) 4- Azidomethyl- 2. 6- difluorobenzonitrile

En blanding av 4-cyano-2,6-difluorbenzyl metansulfonat (1,61 g, 6,51 mmol; se trinn (iv) ovenfor) og natriumazid (0,72 g, 0,0111 mol) i 10 ml vann og 20 ml DMF ble omrørt ved romtemperatur natten over. Det resulterende ble deretter hellet i 200 ml vann og ekstrahert tre ganger med dietyleter. Den samlede eteriske fase ble vasket fem ganger med vann, tørket (Na2S04) og inndampet. En liten prøve ble inndampet for NMR formål og produktet krystallisert. Resten ble inndampet forsiktig men ikke inntil full tørrhet. Utbytte (teoretisk 1,26 g) ble antatt å være nesten kvantitativt basert på NMR og analytisk HPLC. A mixture of 4-cyano-2,6-difluorobenzyl methanesulfonate (1.61 g, 6.51 mmol; see step (iv) above) and sodium azide (0.72 g, 0.0111 mol) in 10 mL of water and 20 ml DMF was stirred at room temperature overnight. The resulting was then poured into 200 ml of water and extracted three times with diethyl ether. The combined ether phase was washed five times with water, dried (Na 2 SO 4 ) and evaporated. A small sample was evaporated for NMR purposes and the product crystallized. The residue was carefully evaporated but not to complete dryness. Yield (theoretical 1.26 g) was estimated to be nearly quantitative based on NMR and analytical HPLC.

i in

'H NMR (400 MHz, CDC13) 8 7,29 (m, 2H), 4,46 (s, 2H) 1 H NMR (400 MHz, CDCl 3 ) δ 7.29 (m, 2H), 4.46 (s, 2H)

(vi) 4- aminometvl- 2. 6- difluorbenzonitril (vi) 4-aminomethyl-2.6-difluorobenzonitrile

Denne reaksjonen ble utført i henhold til metoden beskrevet i J. Chem. Res. ( M) (1992) 3128. Til en suspensjon av 520 mg 10% Pd/C (50% fuktighet) i 20 ml vann ble tilsatt en løsning av natrium-borhydrid (0,834 g, 0,0221 mol) i 20 ml vann. Noe gassutvikling resulterte. 4-Azidometyl-2,6-difluorbenzonitril (1,26 g, 6,49 mmol; se trinn (v) ovenfor) ble oppløst i 50 ml THF og satt til den vandige blandingen på et isbad over 15 min. Blandingen ble omrørt i 4 timer, hvoretter 20 ml 2M HC1 ble tilsatt og blandingen ble filtrert gjennom Celite. Celite ble skyllet med mer vann og den samlede vandige fase ble vasket med EtOAc og deretter gjort alkalisk med 2M NaOH. Eks tr aksjon tre ganger med metylenklorid fulgte og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Utbytte: 0,87 g (80%). This reaction was carried out according to the method described in J. Chem. Res. ( M) (1992) 3128. To a suspension of 520 mg of 10% Pd/C (50% moisture) in 20 ml of water was added a solution of sodium borohydride (0.834 g, 0.0221 mol) in 20 ml of water. Some gas evolution resulted. 4-Azidomethyl-2,6-difluorobenzonitrile (1.26 g, 6.49 mmol; see step (v) above) was dissolved in 50 mL of THF and added to the aqueous mixture on an ice bath over 15 min. The mixture was stirred for 4 hours, after which 20 mL of 2M HCl was added and the mixture was filtered through Celite. Celite was rinsed with more water and the combined aqueous phase was washed with EtOAc and then made alkaline with 2M NaOH. Extraction three times with methylene chloride followed and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 0.87 g (80%).

'H NMR (400 MHz, CDCI3) 8 7,20 (m, 2H), 3,96 (s, 2H), 1,51 (bred, 2H) 1 H NMR (400 MHz, CDCl 3 ) δ 7.20 (m, 2H), 3.96 (s, 2H), 1.51 (broad, 2H)

(vii) 2. 6- difluor- 4- terf- butoksvkarbonvlaminometvlbenzonitril (vii) 2. 6- difluoro- 4- tert-butoxycarbonylaminomethylbenzonitrile

En løsning av 4-aminometyl-2,6-difluorbenzonitril (0,876 g, 5,21 mmol; se trinn (vi) ovenfor) ble oppløst i 50 ml THF og di-te/t-butyl dikarbonat (1,14 g, 5,22 mmol) i 10 ml THF ble tilsatt. Blandingen ble omrørt i 3,5 timer. THF ble inndampet og residuet ble fordelt mellom vann og EtOAc. Det organiske laget ble vasket tre ganger med 0,5 M HC1 og vann, tørket (Na2S04) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 1,38 g (99%). A solution of 4-aminomethyl-2,6-difluorobenzonitrile (0.876 g, 5.21 mmol; see step (vi) above) was dissolved in 50 mL of THF and di-te/t-butyl dicarbonate (1.14 g, 5 .22 mmol) in 10 ml of THF was added. The mixture was stirred for 3.5 hours. The THF was evaporated and the residue partitioned between water and EtOAc. The organic layer was washed three times with 0.5 M HCl and water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 1.38 g (99%).

'H NMR (300 MHz, CDCI3) 8 7,21 (m,2H), 4,95 (bred, 1H), 4,43 (bred, 2H), 1,52 (s, 9H) 1 H NMR (300 MHz, CDCl 3 ) δ 7.21 (m, 2H), 4.95 (broad, 1H), 4.43 (broad, 2H), 1.52 (s, 9H)

(viii) Boe- Pab( 2. 6- diF)( OH) (viii) Boe- Pab( 2. 6- diF)( OH)

En blanding av 2,6-difluor-4-ferf-butoksykarbonylaminometylbenzonitril (1,38 g, 5,16 mmol; se trinn (vii) ovenfor), hydroksylamin-hydroklorid (1,08 g, 0,0155 mol) og trietylamin (1,57 g, 0,0155 mol) i 20 ml etanol ble omrørt ved romtemperatur i 36 timer. Løsningsmidlet ble avdampet og residuet ble fordelt mellom vann og metylenklorid. Det organiske laget ble vasket med vann, tørket (Na2SC«4) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 1,43 g (92%). A mixture of 2,6-difluoro-4-terf-butoxycarbonylaminomethylbenzonitrile (1.38 g, 5.16 mmol; see step (vii) above), hydroxylamine hydrochloride (1.08 g, 0.0155 mol) and triethylamine ( 1.57 g, 0.0155 mol) in 20 ml of ethanol was stirred at room temperature for 36 hours. The solvent was evaporated and the residue was partitioned between water and methylene chloride. The organic layer was washed with water, dried (Na2SO4) and evaporated. The product could be used without further purification. Yield: 1.43 g (92%).

'H NMR (500 MHz, CD3OD) 8 7,14 (m, 2H), 4,97 (bred, 1H), 4,84 (bred, 2H), 4,40 (bred, 2H), 1,43 (s, 9H) 1H NMR (500 MHz, CD3OD) δ 7.14 (m, 2H), 4.97 (broad, 1H), 4.84 (broad, 2H), 4.40 (broad, 2H), 1.43 ( pp. 9H)

(ix) Boc- Pab( 2. 6- diF) x HOAc (ix) Boc-Pab(2.6-diF) x HOAc

Denne reaksjonen ble utført i henhold til metoden beskrevet av Judkins et al, Synth. Comm. This reaction was carried out according to the method described by Judkins et al, Synth. Comm.

(i998) 4351. Boc-Pab(2,6-diF)(OH) (1,32 g, 4,37 mmol; se trinn (viii) ovenfor), eddiksyreanhydrid (0,477 g, 4,68 mmol) og 442 mg 10% Pd/C (50% fuktighet) i 100 ml eddiksyre ble hydrogenen ved 5 atm trykk i 3,5 timer. Blandingen ble filtrert gjennom Celite, skyllet med etanol og inndampet. Residuet ble frysetørket fra acetonitril og vann og noen få dråper etanol. Sub-tittel produktet kunne anvendes uten ytterligere rensning. Utbytte: 0,1,49 g (99%). (i998) 4351. Boc-Pab(2,6-diF)(OH) (1.32 g, 4.37 mmol; see step (viii) above), acetic anhydride (0.477 g, 4.68 mmol) and 442 mg 10% Pd/C (50% humidity) in 100 ml acetic acid was hydrogenated at 5 atm pressure for 3.5 hours. The mixture was filtered through Celite, rinsed with ethanol and evaporated. The residue was freeze-dried from acetonitrile and water and a few drops of ethanol. The sub-title product could be used without further purification. Yield: 0.1.49 g (99%).

'H NMR (400 MHz, CD3OD) 8 7,45 (m, 2H), 4,34 (s, 2H), 1,90 (s, 3H), 1,40 (s, 9H) 1 H NMR (400 MHz, CD 3 OD) δ 7.45 (m, 2H), 4.34 (s, 2H), 1.90 (s, 3H), 1.40 (s, 9H)

(x) Boc- Pab( 2. 6- diF)( Teoc) (x) Boc- Pab( 2. 6- diF)( Teoc)

Til en løsning av Boc-Pab(2,6-diF) x HOAc (1,56 g, 5,49 mmol; se trinn (ix) ovenfor) i 100 ml THF og 1 ml vann ble det tilsatt 2-(trimetylsilyl)etyl p-nitrofenylkarbonat (1,67 g, 5,89 mmol). En løsning av kaliumkarbonat (1,57 g, 0,0114 mol) i 20 ml vann ble tilsatt dråpevis over 5 min. Blandingen ble omrørt natten over. THF ble inndampet og residuet ble fordelt mellom vann og metylenklorid. Det vandige laget ble ekstrahert med metylenklorid og den samlede organiske fase ble vasket to ganger med vandig natriumbikarbonat, tørket (Na2S04) og inndampet. Flash kromatografi på silikagel med heptan/EtOAc = 2/1 ga 1,71 g (73%) av ren forbindelse. To a solution of Boc-Pab(2,6-diF) x HOAc (1.56 g, 5.49 mmol; see step (ix) above) in 100 mL THF and 1 mL water was added 2-(trimethylsilyl) ethyl p-nitrophenyl carbonate (1.67 g, 5.89 mmol). A solution of potassium carbonate (1.57 g, 0.0114 mol) in 20 mL of water was added dropwise over 5 min. The mixture was stirred overnight. The THF was evaporated and the residue was partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride and the combined organic phase was washed twice with aqueous sodium bicarbonate, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with heptane/EtOAc = 2/1 gave 1.71 g (73%) of pure compound.

'H NMR (400 MHz, CDC13) 8 7,43 (m, 2H), 4,97 (bred, 1H), 4,41 (bred, 2H), 4,24 (m, 2H), 1,41 (s, 9H), 1,11 (m, 2H), 0,06 (s, 9H) 1 H NMR (400 MHz, CDCl 3 ) δ 7.43 (m, 2H), 4.97 (broad, 1H), 4.41 (broad, 2H), 4.24 (m, 2H), 1.41 ( s, 9H), 1.11 (m, 2H), 0.06 (s, 9H)

(xi) Boc- Aze- Pab( 2. 6- diF)( Teoc) (xi) Boc- Aze- Pab( 2. 6- diF)( Teoc)

Boc-Pab(2,6-diF)(Teoc) (1,009 g, 2,35 mmol; se trinn (x) ovenfor) ble oppløst i 50 ml EtOAc mettet med HCl(g). Blandingen ble latt stå i 10 min., inndampet og oppløst i 18 ml DMF og deretter avkjølt på et isbad. Boc-Aze-OH (0,450 g, 2,24 mmol), PyBOP (1,24 g, 2,35 mmol) og til slutt ble diisopropyletylamin (1,158 g, 8,96 mmol) tilsatt. Reaksjonsblandingen ble omrørt i 2 timer og deretter hellet i 350 ml vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med saltvann, tørket (Na2S04) og inndampet. Flash kromatografi på silikagel med heptan:EtOAc (1:3) ga 1,097 g (96%) av den ønskede forbindelse. Boc-Pab(2,6-diF)(Teoc) (1.009 g, 2.35 mmol; see step (x) above) was dissolved in 50 mL of EtOAc saturated with HCl(g). The mixture was allowed to stand for 10 min., evaporated and dissolved in 18 ml of DMF and then cooled in an ice bath. Boc-Aze-OH (0.450 g, 2.24 mmol), PyBOP (1.24 g, 2.35 mmol) and finally diisopropylethylamine (1.158 g, 8.96 mmol) were added. The reaction mixture was stirred for 2 h and then poured into 350 mL of water and extracted three times with EtOAc. The combined organic phase was washed with brine, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with heptane:EtOAc (1:3) gave 1.097 g (96%) of the desired compound.

<*>H NMR (500 MHz, CDC13) 8 7,46 (m, 2H), 4,65-4,5 (m, 3H), 4,23 (m, 2H), 3,87 (m, 1H), 3,74 (m, 1H), 2,45-2,3 (m, 2H), 1,40 (s, 9H), 1,10 (m, 2H), 0,05 (s, 9H) <*>H NMR (500 MHz, CDCl 3 ) δ 7.46 (m, 2H), 4.65-4.5 (m, 3H), 4.23 (m, 2H), 3.87 (m, 1H ), 3.74 (m, 1H), 2.45-2.3 (m, 2H), 1.40 (s, 9H), 1.10 (m, 2H), 0.05 (s, 9H)

(xii) Ph( 3- Cl)( 5- OCHF2)- r/ g) CH( OH) C( 0)- Aze- Pab( 2. 6- diF)( Teoc) (xii) Ph( 3- Cl)( 5- OCHF2)- r/ g) CH( OH) C( 0)- Aze- Pab( 2. 6- diF)( Teoc)

Boc-Aze-Pab(2,6-diF)(Teoc) (0,256 g, 0,500 mmol; se trinn (xi) ovenfor) ble oppløst i 20 ml EtOAc mettet med HCl(g). Blandingen ble latt stå i 10 min. og inndampet og oppløst i 5 ml DMF. Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)OH (0,120 g, 0,475 mmol; se Eksempel l(viii) ovenfor), PyBOP (0,263 g, 0,498 mmol) og til slutt diisopropyletylamin (0,245 g, 1,89 mmol ble tilsatt. Reaksjonsblandingen ble omrørt i 2 timer og deretter hellet i 350 ml vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med saltvann, tørket (Na2S04) og inndampet. Flash kromatografi på silikagel med EtOAc ga 0,184 g (60%) av den ønskede sub-tittelforbindelse. Boc-Aze-Pab(2,6-diF)(Teoc) (0.256 g, 0.500 mmol; see step (xi) above) was dissolved in 20 mL EtOAc saturated with HCl(g). The mixture was left to stand for 10 min. and evaporated and dissolved in 5 ml DMF. Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)OH (0.120 g, 0.475 mmol; see Example 1(viii) above), PyBOP (0.263 g, 0.498 mmol) and finally diisopropylethylamine (0.245 g, 1.89 mmol) was added. The reaction mixture was stirred for 2 h and then poured into 350 mL of water and extracted three times with EtOAc. The combined organic phase was washed with brine, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with EtOAc gave 0.184 g (60%) of the desired sub-title compound.

<*>H NMR (400 MHz, CD3OD, blanding av rotamerer) 8 7,55-7,45 (m, 2H), 7,32 (m, 1H, hoved rotamer), 7,27 (m, 1H, mindre rotamer), 7,2-7,1 (m, 2H), 6,90 (t, 1H, hoved rotamer), 6,86 (t, 1H, mindre rotamer), 5,15 (s, lH,hoved rotamer), 5,12 (m, 1H, mindre rotamer), 5,06 (s, 1H, mindre rotamer), 4,72 (m, 1H, hoved rotamer), 4,6-4,45 (m, 2H), 4,30 (m, 1H, hoved rotamer), 4,24 (m, 2H), 4,13 (m, 1H, hoved rotamer), 4,04 (m, 1H, mindre rotamer), 3,95 (m, 1H, mindre rotamer), 2,62 (m, 1H, mindre rotamer), 2,48 (m, 1H, hoved rotamer), 2,22 (m, 1H, hoved rotamer), 2,10 (m, 1H, mindre rotamer), 1,07 (m, 2H), 0,07 (m, 9H) <*>H NMR (400 MHz, CD3OD, mixture of rotamers) δ 7.55-7.45 (m, 2H), 7.32 (m, 1H, major rotamer), 7.27 (m, 1H, minor rotamer), 7.2-7.1 (m, 2H), 6.90 (t, 1H, major rotamer), 6.86 (t, 1H, minor rotamer), 5.15 (s, 1H, major rotamer ), 5.12 (m, 1H, minor rotamer), 5.06 (s, 1H, minor rotamer), 4.72 (m, 1H, major rotamer), 4.6-4.45 (m, 2H) , 4.30 (m, 1H, major rotamer), 4.24 (m, 2H), 4.13 (m, 1H, major rotamer), 4.04 (m, 1H, minor rotamer), 3.95 ( m, 1H, minor rotamer), 2.62 (m, 1H, minor rotamer), 2.48 (m, 1H, major rotamer), 2.22 (m, 1H, major rotamer), 2.10 (m, 1H, minor rotamer), 1.07 (m, 2H), 0.07 (m, 9H)

(xiii) Ph( 3- Cl)( 5- OCHF2)-(/ g) CH( OH) C( Q)- Aze- Pab( 2. 6- diF) (xiii) Ph( 3- Cl)( 5- OCHF2)-(/ g) CH( OH) C( Q)- Aze- Pab( 2. 6- diF)

Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,6-diF)(Teoc) (81 mg, 0,127 mmol; se trinn (xii) ovenfor) ble oppløst i 0,5 ml metylenklorid og avkjølt på et isbad. TFA (3 ml) ble tilsatt og reaksjonen ble latt stå i 75 min. TFA ble inndampet og residuet ble fryse tørket fra vann og acetonitril. Råproduktet ble renset ved preparativ RPLC med CH3CN:0,1M NH4OAC (35:65) for å produsere 39 mg (55%) av tittelforbindelsen som dens HOAc salt, renhet: 99%. Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,6-diF)(Teoc) (81 mg, 0.127 mmol; see step (xii ) above) was dissolved in 0.5 ml methylene chloride and cooled in an ice bath. TFA (3 mL) was added and the reaction was allowed to stand for 75 min. The TFA was evaporated and the residue was freeze dried from water and acetonitrile. The crude product was purified by preparative RPLC with CH3CN:0.1M NH4OAC (35:65) to produce 39 mg (55%) of the title compound as its HOAc salt, purity: 99%.

'H NMR (400 MHz, CD3OD blanding av rotamerer) 8 7,5-7,4 (m, 2H), 7,32 (m, 1H, hoved rotamer), 7,28 (m, 1H, mindre rotamer), 7,2-7,1 (m, 3H) 6,90 (t, 1H, hoved rotamer), 6,86 (t, mindre rotamer), 5,15 (s, 1H, hoved rotamer), 5,14 (m, 1H, mindre rotamer), 5,07 (s, 1H NMR (400 MHz, CD3OD mixture of rotamers) δ 7.5-7.4 (m, 2H), 7.32 (m, 1H, major rotamer), 7.28 (m, 1H, minor rotamer), 7.2-7.1 (m, 3H) 6.90 (t, 1H, major rotamer), 6.86 (t, minor rotamer), 5.15 (s, 1H, major rotamer), 5.14 ( m, 1H, minor rotamer), 5.07 (s,

1H, mindre rotamer), 4,72 (m, 1H, hoved rotamer), 4,65-4,45 (m, 2H), 4,30 (m, 1H, hoved rotamer), 4,16 (m, 1H, hoved rotamer), 4,03 (m, 1H, mindre rotamer), 3,95 (m, 1H, mindre rotamer), 2,63 (m, 1H, mindre rotamer), 2,48 (m, 1H, hoved rotamer), 2,21 (m, 1H, hoved rotamer), 2,07 (m, 1H, mindre rotamer), 1,89 (s, 3H) 1H, minor rotamer), 4.72 (m, 1H, major rotamer), 4.65-4.45 (m, 2H), 4.30 (m, 1H, major rotamer), 4.16 (m, 1H , major rotamer), 4.03 (m, 1H, minor rotamer), 3.95 (m, 1H, minor rotamer), 2.63 (m, 1H, minor rotamer), 2.48 (m, 1H, major rotamer), 2.21 (m, 1H, major rotamer), 2.07 (m, 1H, minor rotamer), 1.89 (s, 3H)

<13>C-NMR (75 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 171,9,171,2,165,0,162,8,160,4 <13>C-NMR (75 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 171,9,171,2,165,0,162,8,160,4

APCI-MS: (M + 1) = 503/505 m/z. APCI-MS: (M + 1) = 503/505 m/z.

Eksempel 40 Example 40

Ph( 3- Cl)( 5- OCHF9)-(/ ?) CH( OH) C( Q)- Aze- Pab( 2. 6- diF)( OMe) Ph( 3- Cl)( 5- OCHF9)-(/ ?) CH( OH) C( Q)- Aze- Pab( 2. 6- diF)( OMe)

(i) Ph( 3- Cl)( 5- OCHF2)-(/ ?JCH( OH) C( 0)- Aze- Pab( 2. 6- diF)( OMe. Teoc) (i) Ph( 3- Cl)( 5- OCHF2)-(/ ?JCH( OH) C( 0)- Aze- Pab( 2. 6- diF)( OMe. Teoc)

En blanding av Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,6-diF)(Teoc) (64 mg, 0,099 mmol; se Eksempel 39(xii) ovenfor) og O-metyl hydroksylamin-hydroklorid (50 mg, 0,60 mmol) i 4 ml acetonitril ble oppvarmet ved 70°C i 3 timer. Løsningsmidlet ble avdampet og residuet ble fordelt mellom vann og EtOAc. Det vandige laget ble ekstrahert to ganger med EtOAc og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 58 mg (87%). A mixture of Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,6-diF)(Teoc) (64 mg, 0.099 mmol; see Example 39(xii) above) and O-methyl hydroxylamine hydrochloride (50 mg, 0.60 mmol) in 4 mL of acetonitrile was heated at 70°C for 3 hours. The solvent was evaporated and the residue was partitioned between water and EtOAc. The aqueous layer was extracted twice with EtOAc and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 58 mg (87%).

'H NMR (400 MHz, CDC13) 8 7,90 (bt, 1H), 7,46 (m, 1H), 7,25-6,95 (m, 5H), 6,51, t, 1H), 4,88 (s, 1H), 4,83 (m, 1H), 4,6-4,5 (m, 2H), 4,4-3,9 (m, 4H), 3,95 (s, 3H), 3,63 (m, 1H), 2,67 (m, 1H), 2,38 (m, 1H), 1,87 (bred, 1H), 0,98 (m, 2H), 0,01, s, 9H) (ii) Ph( 3- Cl)( 5- OCHF7)-( Æ) CH( OH) C( 0)- Aze- Pab( 2. 6- diF)( OMe) Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)-Aze-Pab(2,6-diF)(OMe,Teoc) (58 mg, 0,086 mmol; , se trinn (i) ovenfor) ble oppløst i 3 ml TFA, avkjølt på et isbad og fikk reagere i 2 timer. TFA ble inndampet og residuet oppløst i EtOAc. Det organiske laget ble vasket to ganger med vandig natriumkarbonat og vann, tørket (Na2S04) og inndampet. Residuet ble frysetørket fra vann og acetonitril, hvilket gir 42 mg (92%) av tittelforbindelsen. Renhet: 94%. 1 H NMR (400 MHz, CDCl 3 ) δ 7.90 (bt, 1H), 7.46 (m, 1H), 7.25-6.95 (m, 5H), 6.51, t, 1H), 4.88 (s, 1H), 4.83 (m, 1H), 4.6-4.5 (m, 2H), 4.4-3.9 (m, 4H), 3.95 (s, 3H), 3.63 (m, 1H), 2.67 (m, 1H), 2.38 (m, 1H), 1.87 (broad, 1H), 0.98 (m, 2H), 0, 01, p, 9H) (ii) Ph( 3- Cl)( 5- OCHF7)-( Æ) CH( OH) C( 0)- Aze- Pab( 2. 6- diF)( OMe) Ph(3- Cl)(5-OCHF2)-(/?;CH(OH)C(0)-Aze-Pab(2,6-diF)(OMe,Teoc) (58 mg, 0.086 mmol; , see step (i) above ) was dissolved in 3 mL TFA, cooled in an ice bath and allowed to react for 2 h. The TFA was evaporated and the residue dissolved in EtOAc. The organic layer was washed twice with aqueous sodium carbonate and water, dried (Na 2 SO 4 ) and evaporated. The residue was lyophilized from water and acetonitrile to give 42 mg (92%) of the title compound Purity: 94%.

'H NMR (300 MHz, CDCI3) 8 7,95 (bt, 1H), 7,2-7,1 (m, 4H), 6,99 (m, 1H), 6,52 (t, 1H), 4,88 (s, 1H), 4,85-4,75 (m, 3H), 4,6-4,45 (m, 2H), 4,29 (bred, 1H), 4,09 (m, 1H), 3,89 (s, 3H), 3,69 (m, 1H), 2,64 (m, 1H), 2,38 (m, 1H), 1,85 (bred, 1H) 1 H NMR (300 MHz, CDCl 3 ) δ 7.95 (bt, 1H), 7.2-7.1 (m, 4H), 6.99 (m, 1H), 6.52 (t, 1H), 4.88 (s, 1H), 4.85-4.75 (m, 3H), 4.6-4.45 (m, 2H), 4.29 (wide, 1H), 4.09 (m, 1H), 3.89 (s, 3H), 3.69 (m, 1H), 2.64 (m, 1H), 2.38 (m, 1H), 1.85 (broad, 1H)

<13>C-NMR (100 MHz; CDCI3): (karbonyl og/eller amidinkarbonatomer) 8 172,1, 169,8, 151,9 <13>C-NMR (100 MHz; CDCl3): (carbonyl and/or amidine carbon atoms) 8 172.1, 169.8, 151.9

APCI-MS: (M + 1) = 533/535 m/z APCI-MS: (M + 1) = 533/535 m/z

Eksempel 41 Example 41

Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( 2. 5- diF) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( 2. 5- diF)

(i) 2. 5- difluor- 4r( metvlsulfinvl)( metyltio) metvllbenzonitril (i) 2. 5-difluoro-4r(methylsulfinyl)(methylthio)methylbenzonitrile

(Metylsulfinyl)(metyltio)metan (3,16 g, 0,0255 mol) ble oppløst i 50 ml tørr THF under argon og deretter avkjølt til -78°C. Butyllitium i heksan (16 ml 1,6M, 0,0256 mol) ble tilsatt dråpevis med omrøring. Blandingen ble omrørt i 15 min. I mens ble en løsning av 2,4,5-trifluorbenzonitril (2,0 g; 0,013 mol) i 50 ml tørr THF avkjølt til -78°C under argon (Methylsulfinyl)(methylthio)methane (3.16 g, 0.0255 mol) was dissolved in 50 mL dry THF under argon and then cooled to -78°C. Butyllithium in hexane (16 mL 1.6M, 0.0256 mol) was added dropwise with stirring. The mixture was stirred for 15 min. Meanwhile, a solution of 2,4,5-trifluorobenzonitrile (2.0 g; 0.013 mol) in 50 mL of dry THF was cooled to -78°C under argon

og førstnevnte løsning ble tilsatt gjennom en kanyle til den sistnevnte løsning over en periode på 3-5 min. Etter 30 min, ble kjølebadet fjernet og når reaksjonen hadde nådd romtemperatur ble den hellet i 200 ml vann. THF ble inndampet og det gjenværende vandige lag ble ekstrahert tre ganger med dietyleter. Den samlede eter fase ble vasket med vann, tørket (Na2SC«4) og inndampet. Råproduktet begynte å krystallisere og kunne anvendes som sådanne i neste trinn. Utbytte: 2,8 g (84%). and the former solution was added through a cannula to the latter solution over a period of 3-5 min. After 30 min, the cooling bath was removed and when the reaction had reached room temperature it was poured into 200 ml of water. The THF was evaporated and the remaining aqueous layer was extracted three times with diethyl ether. The combined ether phase was washed with water, dried (Na 2 SC 4 ) and evaporated. The raw product began to crystallize and could be used as such in the next step. Yield: 2.8 g (84%).

'H NMR (500 MHz, CDC13) 5 7,51-7,44 (m, 2H, hoved diastereomer), 7,39 (dd, 1H, mindre diastereomer), 5,00 (s, 1H, mindre diastereomer), 4,92 (s, 1H, hoved diastereomer), 2,59 (s, 3H, mindre diastereomer), 2,56 (s, 1H, hoved diastereomer), 2,46 (s, 1H, mindre diastereomer), 2,40 (s, 1H, hoved diastereomer) 1 H NMR (500 MHz, CDCl 3 ) δ 7.51-7.44 (m, 2H, major diastereomer), 7.39 (dd, 1H, minor diastereomer), 5.00 (s, 1H, minor diastereomer), 4.92 (s, 1H, major diastereomer), 2.59 (s, 3H, minor diastereomer), 2.56 (s, 1H, major diastereomer), 2.46 (s, 1H, minor diastereomer), 2, 40 (s, 1H, major diastereomer)

(ii) 2. 5- difluor- 4- ofrmvlbenzonitril (ii) 2. 5- Difluoro- 4- fluorobenzonitrile

2,5-difluor-4[(metylsulfinyl)(metyltio)metyl]benzonitril (2,8 g, 0,0107 mol; se trinn (i) ovenfor) ble oppløst i 100 ml THF og 6,5 g konsentrert svovelsyre ble tilsatt. Blandingen fikk stå ved romtemperatur i 6 dager og deretter hellet i 500 ml vann. Ekstraksjon tre ganger med dietyleter fulgte og den samlede eteriske fase ble vasket mange ganger med vann, tørket (Na2SC«4) og inndampet. Råproduktet ble flash kromatografert på silikagel ved anvendelse av heptan:EtOAc (8:2). Utbytte: 1,2 g (67%). Stillingen til formylgruppen ble etablert ved anvendelse av C NMR. Karbonsignaler fra fluorerte karbonatomer ved henholdsvis 160,1 og 158,4 var dubletter og ikke kvartetter, som de ville vært hvis formylgruppe hadde vært i 2-stillingen. 2,5-Difluoro-4[(methylsulfinyl)(methylthio)methyl]benzonitrile (2.8 g, 0.0107 mol; see step (i) above) was dissolved in 100 mL of THF and 6.5 g of concentrated sulfuric acid was added . The mixture was allowed to stand at room temperature for 6 days and then poured into 500 ml of water. Extraction three times with diethyl ether followed and the combined ethereal phase was washed several times with water, dried (Na 2 SC 4 ) and evaporated. The crude product was flash chromatographed on silica gel using heptane:EtOAc (8:2). Yield: 1.2 g (67%). The position of the formyl group was established using C NMR. Carbon signals from fluorinated carbons at 160.1 and 158.4, respectively, were doublets and not quartets, as they would have been if the formyl group had been in the 2-position.

'H NMR (300 MHz, CDC13) 8 10,36 (d, 1H), 7,72 (dd, 1H), 7,54 (dd, 1H) 1 H NMR (300 MHz, CDCl 3 ) δ 10.36 (d, 1H), 7.72 (dd, 1H), 7.54 (dd, 1H)

(iii) 2. 5- difluor- 4- hvdroksvmetvlbenzonitril (iii) 2. 5-difluoro-4-hydroxymethylbenzonitrile

2,5-difluor-4-formylbenzonitril (3,60 g, 0,0215 mol; se trinn (ii) ovenfor) ble oppløst i 50 ml metanol og avkjølt på et isbad. Natrium-borhydrid (0,815 g, 0,0215 mol) ble tilsatt i porsjoner med omrøring og reaksjonen ble latt stå i 45 min. Vann (300 ml) ble tilsatt og deretter ble forsiktig 2M HC1 tilsatt inntil en sur pH ble oppnådd. Blandingen ble ekstrahert tre ganger med dietyleter og den samlede eteriske fase ble vasket med vann, tørket (Na2S04) og inndampet. Råproduktet krystallisert fort og kunne anvendes uten ytterligere rensning. Utbytte: 3,1 g (85%). 2,5-Difluoro-4-formylbenzonitrile (3.60 g, 0.0215 mol; see step (ii) above) was dissolved in 50 mL of methanol and cooled in an ice bath. Sodium borohydride (0.815 g, 0.0215 mol) was added in portions with stirring and the reaction was allowed to stand for 45 min. Water (300 mL) was added and then 2M HCl was carefully added until an acidic pH was obtained. The mixture was extracted three times with diethyl ether and the combined ether phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The crude product crystallized quickly and could be used without further purification. Yield: 3.1 g (85%).

'H NMR (300 MHz, CDC13) 8 7,45 (dd, 1H), 7,30 (dd, 1H), 4,85 (s, 2H), 2,10 (bred, 1H) 1 H NMR (300 MHz, CDCl 3 ) δ 7.45 (dd, 1H), 7.30 (dd, 1H), 4.85 (s, 2H), 2.10 (broad, 1H)

(iv) 4- cvano- 2. 5- difluorbenzyl metansulfonat (iv) 4-cvano-2.5-difluorobenzyl methanesulfonate

Til en is avkjølt løsning av 2,5-difluor-4-hydroksymetylbenzonitril (3,10 g, 0,0183 mol; se trinn (iii) ovenfor) og metansulfonylklorid (2,21 g, 0,0192 mol) i 60 ml metylenklorid ble det tilsatt trietylamin (1,95 g, 0,0192 mol) med omrøring. Etter 1,5 timer ved 0°C ble blandingen vasket med vann, tørket (Na2S04) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 4,5 g (99%). To an ice-cooled solution of 2,5-difluoro-4-hydroxymethylbenzonitrile (3.10 g, 0.0183 mol; see step (iii) above) and methanesulfonyl chloride (2.21 g, 0.0192 mol) in 60 mL of methylene chloride triethylamine (1.95 g, 0.0192 mol) was added with stirring. After 1.5 hours at 0°C, the mixture was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 4.5 g (99%).

<!>H NMR (300 MHz, CDCI3) 8 7,45-7,35 (m, 2H), 5,32 (s, 2H), 3,13 (s, 3H) <!>H NMR (300 MHz, CDCl 3 ) δ 7.45-7.35 (m, 2H), 5.32 (s, 2H), 3.13 (s, 3H)

(v) 4- Azidometvl- 2. 5- difluorbenzonitril (v) 4-Azidomethyl-2.5-difluorobenzonitrile

En blanding av 4-cyano-2,5-difluorbenzylmetansulfonat (4,5 g, 0,0182 mol; se trinn (iv) ovenfor) og natriumazid (2,0 g, 0,031 mol) i 20 ml vann og 40 ml DMF ble omrørt ved romtemperatur i 2 timer. Den ble deretter hellet i 300 ml vann og ekstrahert tre ganger med dietyleter. Den samlede eteriske fase ble vasket mange ganger med vann, tørket (Na2SC«4) og inndampet. En liten prøve ble inndampet for NMR formål og produktet krystallisert. Resten ble inndampet forsiktig men ikke til fullstendig tørrhet. Utbytte (teoretisk 3,5 g) er antatt å være nesten kvantitativt basert på NMR og analytisk HPLC. A mixture of 4-cyano-2,5-difluorobenzyl methanesulfonate (4.5 g, 0.0182 mol; see step (iv) above) and sodium azide (2.0 g, 0.031 mol) in 20 mL of water and 40 mL of DMF was stirred at room temperature for 2 hours. It was then poured into 300 ml of water and extracted three times with diethyl ether. The combined ethereal phase was washed several times with water, dried (Na 2 SC 4 ) and evaporated. A small sample was evaporated for NMR purposes and the product crystallized. The residue was carefully evaporated but not to complete dryness. Yield (theoretical 3.5 g) is estimated to be almost quantitative based on NMR and analytical HPLC.

'H NMR (500 MHz, CDCI3) 8 ,38 (dd, 1H), 7,32 (dd, 1H), 4,54 (s, 2H) 1 H NMR (500 MHz, CDCl 3 ) δ .38 (dd, 1H), 7.32 (dd, 1H), 4.54 (s, 2H)

(vi) 4- aminometvl- 2. 5- difluorbenzonitril (vi) 4-aminomethyl-2.5-difluorobenzonitrile

Denne reaksjonen ble utført i henhold til metoden beskrevet i J. Chem. Res. ( M) (1992) 3128. Til en suspensjon av 300 mg 10% Pd/C (50% fuktighet) i 20 ml vann ble det tilsatt en løsning av natrium-borhydrid (0,779 g, 0,0206 mol) i 20 ml vann. Noe gassutvikling resulterte. 4-Azidometyl-2,5-difluorbenzonitril (1,00 g, 5,15 mmol; fra trinn (v) ovenfor) ble oppløst i 60 ml THF og satt til den vandige blandingen på et isbad. Blandingen ble omrørt i 1,5 timer hvoretter 10 ml 2M HC1 ble tilsatt og blandingen ble filtrert gjennom Celite. Celite ble skyllet med mer vann og den samlede vandige fase ble vasket med EtOAc og deretter gjort alkalisk med 2M NaOH. Ekstraksjon tre ganger med metylenklorid fulgte og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Utbytte: 0,47 g (54%). This reaction was carried out according to the method described in J. Chem. Res. ( M) (1992) 3128. To a suspension of 300 mg of 10% Pd/C (50% moisture) in 20 ml of water was added a solution of sodium borohydride (0.779 g, 0.0206 mol) in 20 ml of water . Some gas evolution resulted. 4-Azidomethyl-2,5-difluorobenzonitrile (1.00 g, 5.15 mmol; from step (v) above) was dissolved in 60 mL of THF and added to the aqueous mixture in an ice bath. The mixture was stirred for 1.5 hours after which 10 mL of 2M HCl was added and the mixture was filtered through Celite. Celite was rinsed with more water and the combined aqueous phase was washed with EtOAc and then made alkaline with 2M NaOH. Extraction three times with methylene chloride followed and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Yield: 0.47 g (54%).

'H NMR (300 MHz, CDC13) 6 7,39 (dd, 1H), 7,29 (dd, 1H), 3,99 (s, 2H), 1,45 (bred, 2H) 1 H NMR (300 MHz, CDCl 3 ) δ 7.39 (dd, 1H), 7.29 (dd, 1H), 3.99 (s, 2H), 1.45 (broad, 2H)

(vii) 2. 5- difluor- 4- fe^ butoksvkarbonvlaminometvlbenzonitril (vii) 2. 5- difluoro-4- phenylbutoxycarbonylaminomethylbenzonitrile

En løsning av 4-aminometyl-2,5-difluorbenzonitril (0,46 g, 2,7 mmol; se trinn (vi) ovenfor) og di-tørf-butyl dikarbonat (0,60 g , 2,7 mmol) i 10 ml THF ble omrørt natten over. THF ble inndampet og residuet ble fordelt mellom vann og EtOAc. Det organiske laget ble vasket med vann, tørket (Na2S04) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 0,71 g (97%). A solution of 4-aminomethyl-2,5-difluorobenzonitrile (0.46 g, 2.7 mmol; see step (vi) above) and di-tert-butyl dicarbonate (0.60 g , 2.7 mmol) in 10 ml of THF was stirred overnight. The THF was evaporated and the residue partitioned between water and EtOAc. The organic layer was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 0.71 g (97%).

'H NMR (300 MHz, CDC13) 8 7,35-7,2 (m, 2H), 5,11 (bred triplett, 1H), 4,38 (d, 2H), 1,45 1 H NMR (300 MHz, CDCl 3 ) δ 7.35-7.2 (m, 2H), 5.11 (broad triplet, 1H), 4.38 (d, 2H), 1.45

(s, 9H) (pp, 9H)

(viii) Boc- Pab( 2. 5- diF)( OH) (viii) Boc- Pab( 2. 5- diF)( OH)

En blanding av 2,5-difluor-4-terr-butoksykarbonylaminometylbenzonitril (0,70 g, 2,6 mmol; se trinn (vii) ovenfor), hydroksylamin-hydroklorid (0,54 g, 7,8 mmol) og trietylamin (0,79 g, 7,8 mmol) i 10 ml etanol ble omrørt ved romtemperatur i 6 dager. Den ble deretter fordelt mellom vann og metylenklorid. Det vandige laget ble ekstrahert med metylenklorid og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 0,72 g (92%). A mixture of 2,5-difluoro-4-tert-butoxycarbonylaminomethylbenzonitrile (0.70 g, 2.6 mmol; see step (vii) above), hydroxylamine hydrochloride (0.54 g, 7.8 mmol) and triethylamine ( 0.79 g, 7.8 mmol) in 10 ml of ethanol was stirred at room temperature for 6 days. It was then partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 0.72 g (92%).

<*>H NMR (500 MHz, CD3OD) 8 7,27 (dd, 1H), 7,12 (dd, 1H), 4,29 (s, 2H), 1,47 (s, 9H) <*>H NMR (500 MHz, CD3OD) δ 7.27 (dd, 1H), 7.12 (dd, 1H), 4.29 (s, 2H), 1.47 (s, 9H)

(ix) Boc- Pab( 2. 5- diF) x HOAc (ix) Boc-Pab(2.5-diF) x HOAc

Denne reaksjonen ble utført i henhold til metoden beskrevet av Judkins et al, Synth. Comm. This reaction was carried out according to the method described by Judkins et al, Synth. Comm.

(1998) 4351. Boc-Pab(2,5-diF)(OH) (0,70 g, 2,3 mmol; se trinn (viii) ovenfor), eddiksyreanhydrid (0,25 g, 2,4 mmol) og 230 mg 10% Pd/C (50% fuktighet) i 70 ml eddiksyre ble hydrogenen ved 5 atm trykk i 2,5 timer. Blandingen ble filtrert gjennom Celite og inndampet. Residuet ble fryse tørket fra acetonitril og vann. Produktet kunne anvendes uten ytterligere rensning i neste trinn. Utbytte: 0,80 g (100%). (1998) 4351. Boc-Pab(2,5-diF)(OH) (0.70 g, 2.3 mmol; see step (viii) above), acetic anhydride (0.25 g, 2.4 mmol) and 230 mg of 10% Pd/C (50% humidity) in 70 ml of acetic acid was hydrogenated at 5 atm pressure for 2.5 hours. The mixture was filtered through Celite and evaporated. The residue was freeze dried from acetonitrile and water. The product could be used without further purification in the next step. Yield: 0.80 g (100%).

'H NMR (500 MHz, CD3OD) 8 7,49 (dd, 1H), 7,31 (dd, 1H), 4,33 (s, 2H), 1,91 (s, 3H), 1,46 (s, 9H) 1 H NMR (500 MHz, CD 3 OD) δ 7.49 (dd, 1H), 7.31 (dd, 1H), 4.33 (s, 2H), 1.91 (s, 3H), 1.46 ( pp. 9H)

(x) Boc- Pab( 2. 5- diF)( Teoc) (x) Boc- Pab( 2. 5- diF)( Teoc)

Til en suspensjon av Boc-Pab(2,5-diF) x HOAc (0,80 g, 2,3 mmol; se trinn (ix) ovenfor) i 50 ml THF ble det tilsatt 2-(trimetylsilyl)etyl p-nitrofenylkarbonat (0,85 g, 3,0 mmol). En løsning av kaliumkarbonat (0,80 g, 5,8 mmol) i 10 ml vann ble tilsatt dråpevis. Blandingen ble omrørt natten over. Overskudd av Teoc reagens ble ødelagt ved tilsetning av glycin (0,100 g) og kaliumkarbonat (0,75 g) til løsningen, det ble latt reagere i ytterligere 2 timer. THF ble inndampet og residuet ble fordelt mellom vann og metylenklorid. Det vandige laget ble ekstrahert med metylenklorid og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Flash kromatografi på silikagel med heptan:EtOAc (2:1) ga 0,72 g (72%) av ren forbindelse. To a suspension of Boc-Pab(2,5-diF) x HOAc (0.80 g, 2.3 mmol; see step (ix) above) in 50 mL of THF was added 2-(trimethylsilyl)ethyl p-nitrophenyl carbonate (0.85 g, 3.0 mmol). A solution of potassium carbonate (0.80 g, 5.8 mmol) in 10 mL of water was added dropwise. The mixture was stirred overnight. Excess Teoc reagent was destroyed by adding glycine (0.100 g) and potassium carbonate (0.75 g) to the solution, it was allowed to react for a further 2 hours. The THF was evaporated and the residue was partitioned between water and methylene chloride. The aqueous layer was extracted with methylene chloride and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with heptane:EtOAc (2:1) gave 0.72 g (72%) of pure compound.

<*>H NMR (400 MHz, CDC13) 8 8,00 (dd, 1H), 7,15 (dd, 1H), 4,98 (bred, 1H), 4,36 (bd, 2H), 4,24 (m, 2H), 1,45 (s, 9H), 1,12 (m, 2H), 0,07 (s, 9H) <*>H NMR (400 MHz, CDCl 3 ) δ 8.00 (dd, 1H), 7.15 (dd, 1H), 4.98 (broad, 1H), 4.36 (bd, 2H), 4, 24 (m, 2H), 1.45 (s, 9H), 1.12 (m, 2H), 0.07 (s, 9H)

(xi) H- Pab( 2. 5- diF)( Teoc) x 2 HC1 (xi) H- Pab( 2. 5- diF)( Teoc) x 2 HCl

Boc-Pab(2,5-diF)(Teoc) (0,38 g, 0,88 mmol; se trinn (x) ovenfor) ble oppløst i 50 ml EtOAc mettet med HCl(g). Blandingen ble latt stå i 30 min og inndampet. Boc-Pab(2,5-diF)(Teoc) (0.38 g, 0.88 mmol; see step (x) above) was dissolved in 50 mL EtOAc saturated with HCl(g). The mixture was left for 30 min and evaporated.

•h NMR (500 MHz, CD3OD) 8 7,75-7,6 (m, 2H), 4,46 (m, 2H), 4,3 (s, 2H), 1,15 (m, 2H), 0,07 (s, 9H) h NMR (500 MHz, CD 3 OD) δ 7.75-7.6 (m, 2H), 4.46 (m, 2H), 4.3 (s, 2H), 1.15 (m, 2H), 0.07 (s, 9H)

(xii) Boc- Aze- Pab( 2. 5- diF)( Teoc) (xii) Boc- Aze- Pab( 2. 5- diF)( Teoc)

Til en omrørt løsning av Boc-Aze-OH (0,189 g, 0,94 mmol), H-Pab(2,5-diF)(Teoc) x 2 HC1 (0,36 g, 0,89 mmol; se trinn (xi) ovenfor) og PyBOP (0,54 g, 1,03 mmol) i 5 ml DMF ble tilsatt diisopropyletylamin (0,49 g, 3,8 mmol) og blandingen fikk reagere natten over. Det resulterende ble deretter hellet i vandig natriumbikarbonat og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Flash kromatografi på silikagel med heptan:EtOAc (3:7) ga en tilstrekkelig ren forbindelse. Utbytte: 0,25 g (48%). To a stirred solution of Boc-Aze-OH (0.189 g, 0.94 mmol), H-Pab(2,5-diF)(Teoc) x 2 HCl (0.36 g, 0.89 mmol; see step ( xi) above) and PyBOP (0.54 g, 1.03 mmol) in 5 mL DMF was added diisopropylethylamine (0.49 g, 3.8 mmol) and the mixture was allowed to react overnight. The resulting was then poured into aqueous sodium bicarbonate and extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. Flash chromatography on silica gel with heptane:EtOAc (3:7) gave a sufficiently pure compound. Yield: 0.25 g (48%).

'H NMR (500 MHz, CDC13) 8 7,98 (dd, 1H), 7,13 (dd, 1H), 4,69 (m, 1H), 4,53 (m, 2H), 4,22 (m, 2H), 3,92 (m, 1H), 3,79 (m, 1H), 2,55-2,35 (m, 2H), 1,44 (s, 9H), 1,11 (m, 2H), 0,06 (s, 9H) 1 H NMR (500 MHz, CDCl 3 ) δ 7.98 (dd, 1H), 7.13 (dd, 1H), 4.69 (m, 1H), 4.53 (m, 2H), 4.22 ( m, 2H), 3.92 (m, 1H), 3.79 (m, 1H), 2.55-2.35 (m, 2H), 1.44 (s, 9H), 1.11 (m , 2H), 0.06 (s, 9H)

(xiii) H- Aze- Pab( 2. 5- diF)( Teoc) x 2 HC1 (xiii) H- Aze- Pab( 2. 5- diF)( Teoc) x 2 HC1

Boc-Aze-Pab(2,5-diF)(Teoc) (0,25 g, 0,49 mmol; se trinn (xii) ovenfor) ble oppløst i 50 ml EtOAc mettet med HCl(g). Blandingen ble latt stå i 30 min. og inndampet. Produktet ble anvendt i neste trinn uten ytterligere rensning. Utbytte: 0,23 g (97%). Boc-Aze-Pab(2,5-diF)(Teoc) (0.25 g, 0.49 mmol; see step (xii) above) was dissolved in 50 mL of EtOAc saturated with HCl(g). The mixture was left to stand for 30 min. and evaporated. The product was used in the next step without further purification. Yield: 0.23 g (97%).

'H NMR (400 MHz, CD3OD) 8 7,59 (dd, 1H), 7,47 (dd, 1H), 5,14 (m, 1H), 4,54 (m, 2H), 4,48 (m,2H), 4,15 (m, 1H), 3,96 (m, 1H), 2,87 (m, 1H), 2,56 (m, 1H), 1,17 (m, 2H), 0,05 1 H NMR (400 MHz, CD 3 OD) δ 7.59 (dd, 1H), 7.47 (dd, 1H), 5.14 (m, 1H), 4.54 (m, 2H), 4.48 ( m, 2H), 4.15 (m, 1H), 3.96 (m, 1H), 2.87 (m, 1H), 2.56 (m, 1H), 1.17 (m, 2H), 0.05

(s, 9H) (pp, 9H)

(xiv) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab( 2. 5- diF)( Teoc) (xiv) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab( 2. 5- diF)( Teoc)

Til en løsning av Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)OH (0,12 g, 0,47 mmol; se Eksempel l(viii) ovenfor), H-Aze-Pab(2,5-diF)(Teoc) x 2 HC1 (0,23 g, 0,47 mmol; se trinn (xiii) ovenfor) og PyBOP (0,27 g, 0,52 mmol) i 10 ml DMF ble det tilsatt diisopropyletylamin (0,245 g, 1,90 mmol) og blandingen ble omrørt natten over. Det resulterende ble hellet i vann og ekstrahert tre ganger med EtOAc. Den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Flash kromatografi på silikagel med EtOAc ga 100 mg av en ren fraksjon og 30 mg av en 90% ren fraksjon. Total utbytte: 0,13 g (41%). To a solution of Ph(3-Cl)(5-OCHF2)-(/?;CH(OH)C(0)OH (0.12 g, 0.47 mmol; see Example 1(viii) above), H -Aze-Pab(2,5-diF)(Teoc) x 2 HCl (0.23 g, 0.47 mmol; see step (xiii) above) and PyBOP (0.27 g, 0.52 mmol) in 10 ml DMF was added diisopropylethylamine (0.245 g, 1.90 mmol) and the mixture was stirred overnight. The resulting was poured into water and extracted three times with EtOAc. The combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated .Flash chromatography on silica gel with EtOAc gave 100 mg of a pure fraction and 30 mg of a 90% pure fraction.Total yield: 0.13 g (41%).

'H NMR (400 MHz, CDC13) 8 9,80 (bred, 1H), 8,05 (bt, 1H), 7,94 (dd, 1H),7,20 (m, 1H), 7,2-7,1 (m, 2H), 7,02 (m, 1H), 6,54 (t, 1H), 4,93 (s, 1H), 4,91 (m, 1H), 4,51 (m, 2H), 4,28 (bred, 1H), 4,23 (m, 2H), 4,13 (m, 1H), 3,74 (m, 1H), 2,69 (m, 1H), 2,43 (m, 1H), 1,73 (bred, 1H), 1,11 (m, 2H), 1,11 (s, 9H) 1 H NMR (400 MHz, CDCl 3 ) δ 9.80 (broad, 1H), 8.05 (bt, 1H), 7.94 (dd, 1H), 7.20 (m, 1H), 7.2- 7.1 (m, 2H), 7.02 (m, 1H), 6.54 (t, 1H), 4.93 (s, 1H), 4.91 (m, 1H), 4.51 (m , 2H), 4.28 (broad, 1H), 4.23 (m, 2H), 4.13 (m, 1H), 3.74 (m, 1H), 2.69 (m, 1H), 2 .43 (m, 1H), 1.73 (wide, 1H), 1.11 (m, 2H), 1.11 (s, 9H)

(xv) Ph( 3- Cl)( 5- OCHF?)-( fl) CH( OH) C( Q)- Aze- Pab( 2. 5- diF) (xv) Ph( 3- Cl)( 5- OCHF?)-( fl) CH( OH) C( Q)- Aze- Pab( 2. 5- diF)

Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,5-diF)(Teoc) (60 mg (0,093 mmol) ren fraksjon fra trinn (xiv) ovenfor) ble oppløst i 3 ml TFA og latt stå ved romtemperatur i 1 time. TFA ble inndampet og residuet ble frysetørket fra vann og acetonitril for å produsere 55 mg (96%) av tittelforbindelsen som dens TFA salt, renhet: >99%. Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,5-diF)(Teoc) (60 mg (0.093 mmol) pure fraction from step (xiv) above) was dissolved in 3 mL of TFA and left at room temperature for 1 hour. The TFA was evaporated and the residue was freeze-dried from water and acetonitrile to produce 55 mg (96%) of the title compound as its TFA salt, purity: >99%.

'H NMR (500 MHz, CD3OD blanding av rotamerer) 8 7,55-7,3 (m, 3H), 7,2-7,1 (m, 2H), 6,88 (t, 1H, hoved rotamer), 6,86 (t, 1H, mindre rotamer), 5,22 (m, 1H, mindre rotamer), 1H NMR (500 MHz, CD3OD mixture of rotamers) δ 7.55-7.3 (m, 3H), 7.2-7.1 (m, 2H), 6.88 (t, 1H, main rotamer) , 6.86 (t, 1H, minor rotamer), 5.22 (m, 1H, minor rotamer),

5,20 (s, 1H, hoved rotamer), 5,13 (s, 1H, mindre rotamer), 4,80 (m, 1H, hoved rotamer), 4,6-4,45 (m, 2H), 4,36 (m, 1H, hoved rotamer), 4,19 (m, 1H, hoved rotamer), 4,07 (m, 1H, mindre rotamer), 3,98 (m, 1H, mindre rotamer), 2,70 (m, 1H, mindre rotamer), 2,54 (m, 1H, hoved rotamer), 2,28 (m, 1H, hoved rotamer), 2,14 (m, 1H, mindre rotamer) <13>C-NMR (75 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, blanding av rotamerer) 8 173,0,172,6,172,1,172,0,162,4 5.20 (s, 1H, major rotamer), 5.13 (s, 1H, minor rotamer), 4.80 (m, 1H, major rotamer), 4.6-4.45 (m, 2H), 4 .36 (m, 1H, major rotamer), 4.19 (m, 1H, major rotamer), 4.07 (m, 1H, minor rotamer), 3.98 (m, 1H, minor rotamer), 2.70 (m, 1H, minor rotamer), 2.54 (m, 1H, major rotamer), 2.28 (m, 1H, major rotamer), 2.14 (m, 1H, minor rotamer) <13>C-NMR (75 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, mixture of rotamers) 8 173,0,172,6,172,1,172,0,162,4

APCI-MS: (M + 1) = 503/505 m/z. APCI-MS: (M + 1) = 503/505 m/z.

Eksempel 42 Example 42

Ph( 3- Cl)( 5- OCHF7)-(/ ?) CH( OH) C( 0)- Aze- Pab( 2. 5- diF)( OMe) Ph( 3- Cl)( 5- OCHF7)-(/ ?) CH( OH) C( 0)- Aze- Pab( 2. 5- diF)( OMe)

(i) Ph( 3- Cl)( 5- OCHF7)- r/ g) CH( OH) C( Q)- AzePab( 2. 5- diF)( OMe. Teoc) (i) Ph( 3- Cl)( 5- OCHF7)- r/ g) CH( OH) C( Q)- AzePab( 2. 5- diF)( OMe. Teoc)

En blanding av Ph(3-Cl)(5-OCHF2)-WCH(OH)C(0)-Aze-Pab(2,5-diF)(Teoc) (40 mg, 0,062 mmol; se Eksempel 41(xiv) ovenfor) og O-metyl hydroksylamin-hydroklorid (58 mg, 0,70 mmol) i 5 ml acetonitril ble oppvarmet ved 70°C i 2 timer. Løsningsmidlet ble avdampet og residuet ble fordelt mellom vann og EtOAc. Det vandige laget ble ekstrahert med EtOAc og den samlede organiske fase ble vasket med vann, tørket (Na2S04) og inndampet. Produktet kunne anvendes uten ytterligere rensning. Utbytte: 35 mg (84%). A mixture of Ph(3-Cl)(5-OCHF2)-WCH(OH)C(0)-Aze-Pab(2,5-diF)(Teoc) (40 mg, 0.062 mmol; see Example 41(xiv) above) and O-methyl hydroxylamine hydrochloride (58 mg, 0.70 mmol) in 5 mL of acetonitrile was heated at 70°C for 2 hours. The solvent was evaporated and the residue was partitioned between water and EtOAc. The aqueous layer was extracted with EtOAc and the combined organic phase was washed with water, dried (Na 2 SO 4 ) and evaporated. The product could be used without further purification. Yield: 35 mg (84%).

<*>H NMR (600 MHz, CDC13) 8 7,99 (bt, 1H), 7,72 (s, 1H), 7,20 (m, 1H) 7,15-7,1 (m, 1H), 7,07 (dd, 1H), 7,01 (m, 1H), 6,53 (t, 1H), 4,90 (s, 1H), 4,88 m, 1H), 4,48 (m, 2H), 4,2-4,1 (m, 3H), 3,95 (s, 3H), 3,67 (m, 1H), 2,68 (m, 1H), 2,41 (m, 1H), 0,97 (m, 2H), 0,07 (s, 9H) (ii) Ph( 3- Cn( 5- OCHF2Vr^ JCH( OH') C( 0)- Aze- Pabr2. 5- diF) rOMe) Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,5-diF)(OMe,Teoc) (35 mg, 0,052 mmol; se trinn (i) ovenfor) ble oppløst i 3 ml TFA og fikk reagere i 30 min. TFA ble inndampet og residuet frysetørket fra vann og acetonitril, hvilket gir 29 mg (99%) av tittelforbindelsen. Renhet: 97%. <*>H NMR (600 MHz, CDCl 3 ) δ 7.99 (bt, 1H), 7.72 (s, 1H), 7.20 (m, 1H) 7.15-7.1 (m, 1H) , 7.07 (dd, 1H), 7.01 (m, 1H), 6.53 (t, 1H), 4.90 (s, 1H), 4.88 m, 1H), 4.48 (m , 2H), 4.2-4.1 (m, 3H), 3.95 (s, 3H), 3.67 (m, 1H), 2.68 (m, 1H), 2.41 (m, 1H), 0.97 (m, 2H), 0.07 (s, 9H) (ii) Ph( 3- Cn( 5- OCHF2Vr^ JCH( OH') C( 0)- Aze- Pabr2. 5- diF ) rOMe) Ph(3-Cl)(5-OCHF2)-(/?>)CH(OH)C(0)-Aze-Pab(2,5-diF)(OMe,Teoc) (35 mg, 0.052 mmol ; see step (i) above) was dissolved in 3 ml of TFA and allowed to react for 30 min. The TFA was evaporated and the residue freeze-dried from water and acetonitrile to give 29 mg (99%) of the title compound. Purity: 97%.

<!>H NMR (300 MHz, CDC13) 8 8,01 (bt, 1H), 7,45 (dd, 1H), 7,20 (m, 1H), 7,15 (m, 1H), 7,09 (dd, 1H), 7,02 (m, 1H), 6,54 (t, 1H), 5,2-5,0 (m, 2H), 4,95-4,85 (m, 2H), 4,6-4,4 (m, 2H), 4,25 (bred, 1H), 4,13 (m, 1H), 3,90 (s, 3H), 3,71 (m, 1H), 2,69 (m, 1H), 2,43 (m, 1H) <13>C-NMR (75 MHz; CDC13): (karbonyl og/eller amidinkarbonatomer) 8 173,0,170,9, 152,6 <!>H NMR (300 MHz, CDCl 3 ) δ 8.01 (bt, 1H), 7.45 (dd, 1H), 7.20 (m, 1H), 7.15 (m, 1H), 7, 09 (dd, 1H), 7.02 (m, 1H), 6.54 (t, 1H), 5.2-5.0 (m, 2H), 4.95-4.85 (m, 2H) , 4.6-4.4 (m, 2H), 4.25 (broad, 1H), 4.13 (m, 1H), 3.90 (s, 3H), 3.71 (m, 1H), 2.69 (m, 1H), 2.43 (m, 1H) <13>C-NMR (75 MHz; CDCl3): (carbonyl and/or amidine carbon atoms) 8 173.0, 170.9, 152.6

APCI-MS: (M + 1) = 533/535 m/z. APCI-MS: (M + 1) = 533/535 m/z.

Eksempel 43 Example 43

Ph( 3- Cl)( 5- OCHFz)-(/ g) CH( OmC( Q)- Aze- Pab( OEt) Ph( 3- Cl)( 5- OCHFz)-(/ g) CH( OmC( Q)- Aze- Pab( OEt)

(i) Ph( 3- Cl)( 5- OCHF9)-(/ ?) CH( OH) C( Q)- Aze- Pab( OEt. Teoc) (i) Ph( 3- Cl)( 5- OCHF9)-(/ ?) CH( OH) C( Q)- Aze- Pab( OEt. Teoc)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (55 mg, 0,090 mmol; se Eksempel l(ix) ovenfor) og O-etylhydroksyl amin-hydroklorid (53 mg, 0,54 mmol) ble oppløst i 4 ml THF. Blandingen ble omrørt ved 60°C i 5 timer. Løsningsmidlet ble avdampet. Residuet ble kromatografert på silikagel, under eluering med metylenklorid:metanol (95:5), hvilket gir 55 mg (93%) av sub-tittelforbindelse. Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (55 mg, 0.090 mmol; see Example 1(ix) above) and O-ethylhydroxyl Amine hydrochloride (53 mg, 0.54 mmol) was dissolved in 4 mL of THF. The mixture was stirred at 60°C for 5 hours. The solvent was evaporated. The residue was chromatographed on silica gel, eluting with methylene chloride:methanol (95:5), yielding 55 mg (93%) of sub-title compound.

'H-NMR (400 MHz; CDCI3): 8 7,84 (bt, 1H), 7,59 (bs, 1H), 7,47 (bd, 1H), 7,29 (bd, 1H), 7,21 (m, 1H), 7,14 (m, 1H), 7,02 (m, 1H), 6,53 (t, 1H), 4,90 (s, 1H), 4,86 (m, 1H), 4,55-4,4 (m, 2H), 4,25-4,1 (m, 5H), 3,69 (m, 1H), 2,66 (m, 1H), 2,41 (m, 1H), 1,33 (t, 3H), 0,98 (m, 2H), 0,02 (s, 9H) 1H-NMR (400 MHz; CDCl 3 ): δ 7.84 (bt, 1H), 7.59 (bs, 1H), 7.47 (bd, 1H), 7.29 (bd, 1H), 7, 21 (m, 1H), 7.14 (m, 1H), 7.02 (m, 1H), 6.53 (t, 1H), 4.90 (s, 1H), 4.86 (m, 1H ), 4.55-4.4 (m, 2H), 4.25-4.1 (m, 5H), 3.69 (m, 1H), 2.66 (m, 1H), 2.41 ( m, 1H), 1.33 (t, 3H), 0.98 (m, 2H), 0.02 (s, 9H)

(ii) Ph( 3- Cl. 5- OCHFz)-(/ ?) CH( OH) C( Q)- Aze- Pab( OEt) (ii) Ph( 3- Cl. 5- OCHFz)-(/ ?) CH( OH) C( Q)- Aze- Pab( OEt)

Til en is-kald løsning av Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OEt, Teoc) (55 mg, 0,084 mmol; se trinn (i) ovenfor) i 0,5 ml metylenklorid ble det tilsatt 3 ml TFA. Blandingen ble omrørt (is-bad) i 160 minutter. Materialet ble renset ved anvendelse av preparativ HPLC. Fraksjonene av interesse ble samlet og frysetørket (2x), hvilket gir 20 mg (47%) av tittelforbindelsen. To an ice-cold solution of Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OEt, Teoc) (55 mg, 0.084 mmol; see step ( i) above) in 0.5 ml methylene chloride was added 3 ml TFA. The mixture was stirred (ice bath) for 160 minutes. The material was purified using preparative HPLC. The fractions of interest were pooled and freeze-dried (2x), yielding 20 mg (47%) of the title compound.

'H-NMR (400 MHz; CD3OD) rotamerer: 8 7,59 (bd, 2H), 7,35 (m, 1H), 7,32 (bd, 2H), 7,25-7,1 (m, 2H), 6,89 (t, 1H, hoved rotamer), 6,86 (t, 1H, mindre rotamer), 5,18 (s, 1H, hoved rotamer), 5,18 (m, 1H, mindre rotamer), 5,11 (s, 1H, mindre rotamer), 4,77 (m, 1H), 4,5-4,3 (m, 3H), 4,2-3,9 (m, 3H), 2,67 (m, 1H, mindre rotamer), 2,52 (m, 1H, hoved rotamer), 2,28 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer), 1,28 (t, 3H) <13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 172,4,171,9,171,4,153,8,152,3 1H-NMR (400 MHz; CD 3 OD) rotamers: δ 7.59 (bd, 2H), 7.35 (m, 1H), 7.32 (bd, 2H), 7.25-7.1 (m, 2H), 6.89 (t, 1H, major rotamer), 6.86 (t, 1H, minor rotamer), 5.18 (s, 1H, major rotamer), 5.18 (m, 1H, minor rotamer) , 5.11 (s, 1H, minor rotamer), 4.77 (m, 1H), 4.5-4.3 (m, 3H), 4.2-3.9 (m, 3H), 2, 67 (m, 1H, minor rotamer), 2.52 (m, 1H, major rotamer), 2.28 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer), 1.28 ( t, 3H) <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 172,4,171,9,171,4,153,8,152,3

MS (m/z) 509 (M - 1)\ 511 (M + 1)<+>MS (m/z) 509 (M - 1)\ 511 (M + 1)<+>

Eksempel 44 Example 44

Ph( 3- Cl)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( OnPr) Ph( 3- Cl)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( OnPr)

(i) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OnPr. Teoc) (i) Ph( 3- Cl)( 5- OCHF2)-(/ ?) CH( OH) C( 0)- Aze- Pab( OnPr. Teoc)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (53 mg, 0,087 mmol; se Eksempel l(ix) ovenfor) og O-n-propylhydroksyl amin-hydroklorid, 58 mg (0,52 mmol) ble oppløst i 4 ml THF. Blandingen ble omrørt ved 60°C i 5 timer. Løsningsmidlet ble avdampet. Residuet ble kromatografert på silikagel, under eluering med metylenklorid:metanol (95:5), hvilket gir 51 mg (88%) av sub-tittelforbindelse. Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (53 mg, 0.087 mmol; see Example 1(ix) above) and O-n-propylhydroxyl amine hydrochloride, 58 mg (0.52 mmol) was dissolved in 4 mL of THF. The mixture was stirred at 60°C for 5 hours. The solvent was evaporated. The residue was chromatographed on silica gel, eluting with methylene chloride:methanol (95:5), yielding 51 mg (88%) of sub-title compound.

'H-NMR (400 MHz; CDC13): 8 7,84 (m, 1H), 7,59 (bs, 1H), 7,47 (bd, 2H), 7,28 (bd, 2H), 7.21 (m, 1H), 7,14 (m, 1H), 7,02 (m, 1H), 6,53 (t, 1H), 4,90 (s, 1H), 4,85 (m, 1H), 4,55-4,4 (m, 2H), 4,2-4,05 (m, 5H), 3,69 (m, 1H), 2,65 (m, 1H), 2,41 (m, 1H), 1,74 (m, 2H), 1,05-0,95 (m, 5H), 0,03 (s, 9H) 1H-NMR (400 MHz; CDCl 3 ): δ 7.84 (m, 1H), 7.59 (bs, 1H), 7.47 (bd, 2H), 7.28 (bd, 2H), 7.21 ( m, 1H), 7.14 (m, 1H), 7.02 (m, 1H), 6.53 (t, 1H), 4.90 (s, 1H), 4.85 (m, 1H), 4.55-4.4 (m, 2H), 4.2-4.05 (m, 5H), 3.69 (m, 1H), 2.65 (m, 1H), 2.41 (m, 1H), 1.74 (m, 2H), 1.05-0.95 (m, 5H), 0.03 (s, 9H)

(ii) Ph( 3- Cl)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( OnPr) (ii) Ph( 3- Cl)( 5- OCHF2)-(/ g) CH( OH) C( 0)- Aze- Pab( OnPr)

Til en is-kald løsning av Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OnPr, Teoc) (51 mg, 0,078 mmol; se trinn (i) ovenfor) i 0,5 ml metylenklorid ble det tilsatt 3 ml TFA. Blandingen ble omrørt (is-bad) i 110 minutter. Materialet ble renset ved anvendelse av preparativ HPLC. Fraksjonen av interesse ble inndampet og frysetørket, hvilket gir 20 mg (47%) av tittelforbindelsen. To an ice-cold solution of Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OnPr, Teoc) (51 mg, 0.078 mmol; see step ( i) above) in 0.5 ml methylene chloride was added 3 ml TFA. The mixture was stirred (ice bath) for 110 minutes. The material was purified using preparative HPLC. The fraction of interest was evaporated and freeze-dried, yielding 20 mg (47%) of the title compound.

'H-NMR (500 MHz; CD3OD) rotamerer: 8 7,61 (bd, 2H), 7,38 (m, 1H), 7,35 (bd, 2H), 7.22 (m, 1H, hoved rotamer), 7,18 (m, 1H), 7,15 (m, 1H, mindre rotamer), 6,92 (t, 1H, 1H-NMR (500 MHz; CD3OD) rotamer: δ 7.61 (bd, 2H), 7.38 (m, 1H), 7.35 (bd, 2H), 7.22 (m, 1H, main rotamer), 7.18 (m, 1H), 7.15 (m, 1H, minor rotamer), 6.92 (t, 1H,

hoved rotamer), 6,89 (t, 1H, mindre rotamer), 5,20 (s, 1H, hoved rotamer), 5,20 (m, 1H, mindre rotamer), 4,80 (m, 1H, hoved rotamer), 4,5-4,4 (m, 2H, omfattende mindre rotamer svarende til hoved ved 4,37), 4,37 (m, 1H, hoved rotamer), 4,18 (m, 1H, hoved rotamer), 4,09 (m, 1H, mindre rotamer), 3,99 (m, 2H), 2,70 (m, 1H, mindre rotamer), 2,54 (m, 1H, hoved rotamer), 2,30 (m, 1H, hoved rotamer), 2,18 (m, 1H, mindre rotamer), 1,73 (m, 2H), 1,01 (t, 3H) major rotamer), 6.89 (t, 1H, minor rotamer), 5.20 (s, 1H, major rotamer), 5.20 (m, 1H, minor rotamer), 4.80 (m, 1H, major rotamer ), 4.5-4.4 (m, 2H, comprising minor rotamer corresponding to main at 4.37), 4.37 (m, 1H, main rotamer), 4.18 (m, 1H, main rotamer), 4.09 (m, 1H, minor rotamer), 3.99 (m, 2H), 2.70 (m, 1H, minor rotamer), 2.54 (m, 1H, major rotamer), 2.30 (m , 1H, major rotamer), 2.18 (m, 1H, minor rotamer), 1.73 (m, 2H), 1.01 (t, 3H)

<13>C-NMR (125 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 171,4,153,8,152,3 <13>C-NMR (125 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 171,4,153,8,152,3

MS (m/z) 523 (M - 1)\ 525 (M + 1)<+>MS (m/z) 523 (M - 1)\ 525 (M + 1)<+>

Eksempel 45 Example 45

Ph( 3- Cl)( 5- OCHF7)-( i?) CH( OH) C( 0)- Aze- Pab( 0/ Pr) Ph( 3- Cl)( 5- OCHF7)-( i?) CH( OH) C( 0)- Aze- Pab( 0/ Pr)

(i) Ph( 3- Cl)( 5- OCHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( OiPr. Teoc) (i) Ph( 3- Cl)( 5- OCHF9)-(/ g) CH( OH) C( Q)- Aze- Pab( OiPr. Teoc)

Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (50 mg, 0,082 mmol; se Eksempel l(ix) ovenfor) og O-i-propylhydroksyl amin-hydroklorid, 55 mg (0,49 mmol) ble oppløst i 4 ml THF. Blandingen ble omrørt ved 60°C i 5 timer. Løsningsmidlet ble avdampet. Residuet ble kromatografert på silikagel, under eluering med metylenkloird:metanol (95:5), hvilket gir 46 mg (84%) av sub-tittelforbindelse. Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(Teoc) (50 mg, 0.082 mmol; see Example 1(ix) above) and O-i-propylhydroxyl amine hydrochloride, 55 mg (0.49 mmol) was dissolved in 4 mL of THF. The mixture was stirred at 60°C for 5 hours. The solvent was evaporated. The residue was chromatographed on silica gel, eluting with methylene chloride:methanol (95:5), yielding 46 mg (84%) of sub-title compound.

'H-NMR (400 MHz; CDC13): 8 7,84 (m, 1H), 7,57 (bs, 1H), 7,48 (bd, 2H), 7,29 (bd, 2H), 7,21 (m, 1H), 7,14 (m, 1H), 7,02 (m,lH), 6,53 (t, 1H), 4,91 (s, 1H), 4,87 (m, 1H), 4,55-4,45 (m, 2H), 4,42 (m, 1H), 4,2-4,1 (m, 3H), 3,69 (m, 1H), 2,66 (m, 1H), 2,42 (m, 1H), 1,30 (d, 6H), 0,98 (m, 2H), 0,02 (s, 9H) 1H-NMR (400 MHz; CDCl 3 ): δ 7.84 (m, 1H), 7.57 (bs, 1H), 7.48 (bd, 2H), 7.29 (bd, 2H), 7, 21 (m, 1H), 7.14 (m, 1H), 7.02 (m,lH), 6.53 (t, 1H), 4.91 (s, 1H), 4.87 (m, 1H ), 4.55-4.45 (m, 2H), 4.42 (m, 1H), 4.2-4.1 (m, 3H), 3.69 (m, 1H), 2.66 ( m, 1H), 2.42 (m, 1H), 1.30 (d, 6H), 0.98 (m, 2H), 0.02 (s, 9H)

(ii) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab( OtPr) (ii) Ph( 3- Cl)( 5- OCHF7)-(/ g) CH( OH) C( Q)- Aze- Pab( OtPr)

Til en is-kald løsning av Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OiPr, Teoc) (46 mg, 0,069 mmol; se trinn (i) ovenfor) i 0,5 ml metylenklorid ble det tilsatt 3 ml TFA. Blandingen ble omrørt (is-bad) i 150 minutter. Materialet ble renset ved anvendelse av preparativ HPLC. Fraksjonen av interesse ble inndampet og frysetørket (2x), hvilket gir 22 mg (58%) av tittelforbindelsen. To an ice-cold solution of Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OiPr, Teoc) (46 mg, 0.069 mmol; see step ( i) above) in 0.5 ml methylene chloride was added 3 ml TFA. The mixture was stirred (ice bath) for 150 minutes. The material was purified using preparative HPLC. The fraction of interest was evaporated and freeze-dried (2x), yielding 22 mg (58%) of the title compound.

'H-NMR (400 MHz; CD3OD) rotamerer: 8 7,59 (d, 2H), 7,35 (m, 1H), 7,32 (d, 2H), 7,19 (m, 1H, hoved rotamer), 7,15 (m, 1H), 7,12 (m, 1H, mindre rotamer), 6,89 (t, 1H, hoved rotamer), 6,86 (t, 1H, mindre rotamer), 5,18 (s, 1H, hoved rotamer), 5,18 (m, 1H, mindre rotamer), 5,12 (s, 1H, mindre rotamer), 4,78 (m, 1H, hoved rotamer), 4,5-3,9 (m, 5H), 2,67 (m, 1H, mindre rotamer), 2,52 (m, 1H, hoved rotamer), 2,28 (m, 1H, hoved rotamer), 2,15 (m, 1H, mindre rotamer), 1,26 (d, 6H) 'H-NMR (400 MHz; CD3OD) rotamers: δ 7.59 (d, 2H), 7.35 (m, 1H), 7.32 (d, 2H), 7.19 (m, 1H, major rotamer ), 7.15 (m, 1H), 7.12 (m, 1H, minor rotamer), 6.89 (t, 1H, major rotamer), 6.86 (t, 1H, minor rotamer), 5.18 (s, 1H, major rotamer), 5.18 (m, 1H, minor rotamer), 5.12 (s, 1H, minor rotamer), 4.78 (m, 1H, major rotamer), 4.5-3 .9 (m, 5H), 2.67 (m, 1H, minor rotamer), 2.52 (m, 1H, major rotamer), 2.28 (m, 1H, major rotamer), 2.15 (m, 1H, minor rotamer), 1.26 (d, 6H)

<13>C-NMR (100 MHz; CD3OD): (karbonyl og/eller amidinkarbonatomer, rotamerer) 8 171,9,171,4,153,6. <13>C-NMR (100 MHz; CD3OD): (carbonyl and/or amidine carbon atoms, rotamers) 8 171,9,171,4,153,6.

MS (m/z) 523 (M - 1)", 525 (M + 1)<+>MS (m/z) 523 (M - 1)", 525 (M + 1)<+>

Eksempel 46 Example 46

Tittelforbindelsen i Eksemplene 3,6, 9,10,13 til 15,17,19,21, 23,25,27, 28, 32,34, 36, 38, 39 og 41 ble testet i Test A ovenfor og ble funnet å vise IC50TT verdier på mindre enn 3,5 uM. De i Eksemplene 3,6,9,10,13,15,17,19,21,23, 27, 32, 34 og 39 ble funnet å vise verdier som var mindre enn 0,02 uM; de i Eksemplene 25 og 28 mindre enn 0,03 \ iM, de i Eksempel 14 mindre enn 0,04 |iM; og de i Eksemplene 38 og 41 mindre enn 0,15 uM. The title compound in Examples 3,6, 9,10,13 to 15,17,19,21, 23,25,27, 28, 32,34, 36, 38, 39 and 41 was tested in Test A above and found to show IC50TT values of less than 3.5 µM. Those in Examples 3, 6, 9, 10, 13, 15, 17, 19, 21, 23, 27, 32, 34 and 39 were found to show values less than 0.02 µM; those in Examples 25 and 28 less than 0.03 µM, those in Example 14 less than 0.04 µM; and those in Examples 38 and 41 less than 0.15 µM.

Eksempel 47 Example 47

. Tittelforbindelsen i Eksemplene 3,6,13,15,17,19,21,23, 25,27, 28, 32 og 34 ble testet i Test D ovenfor og ble funnet å vise en IC50 APTT verdi på mindre enn 1 TM. . The title compound of Examples 3,6,13,15,17,19,21,23, 25,27, 28, 32 and 34 was tested in Test D above and was found to exhibit an IC50 APTT value of less than 1 TM.

Eksempel 48 Example 48

Tittelforbindelsene i Eksemplene 1, 2,4, 5,7,12,16,18,20,22, 24,26,29, 30, 33 og 43 til 45 ble testet i Test E ovenfor og ble funnet å vise oral og/eller parenteral biotilgjengelighet i rotte som den tilsvarende aktive inhibitor (fri amidin). The title compounds of Examples 1, 2,4, 5,7,12,16,18,20,22, 24,26,29, 30, 33 and 43 to 45 were tested in Test E above and were found to exhibit oral and/or or parenteral bioavailability in the rat as the corresponding active inhibitor (free amidine).

Eksempel 49 Example 49

Tittelforbindelsene i Eksemplene 1,2,7, 8,11,12,16,18,20,22, 24,26,29, 33, 37,40,43 og 45 ble testet i Test G ovenfor og ble funnet å bli omdannet til den tilsvarende aktive inhibitor (fri amidin) i levermikrosomer fra mennesker og fra rotter. The title compounds of Examples 1,2,7, 8,11,12,16,18,20,22, 24,26,29, 33, 37,40,43 and 45 were tested in Test G above and found to be converted to the corresponding active inhibitor (free amidine) in human and rat liver microsomes.

Forkortelser Abbreviations

Ac = acetyl Ac = acetyl

AcOH = eddiksyre AcOH = acetic acid

APCI = atmosfærisk trykk kjemisk ionisering (i relasjon til MS) APCI = atmospheric pressure chemical ionization (in relation to MS)

API = atmosfærisk trykk ionisering (i relasjon til MS) API = atmospheric pressure ionization (relative to MS)

vandig = vandig watery = watery

AUC = areal under kurven AUC = area under the curve

Aze = (S)-azetidin-2-karboksylat (hvis ikke på annen måte spesifisert) AzeOH = azetidin-2-karboksylsyre Aze = (S)-azetidine-2-carboxylate (unless otherwise specified) AzeOH = azetidine-2-carboxylic acid

Bn = benzyl Bn = benzyl

Boe = tørf-butyloksykarbonyl Boe = peat-butyloxycarbonyl

BSA = bovint serumalbumin BSA = bovine serum albumin

Bu = butyl Bu = butyl

Bzl = benzyl Bzl = benzyl

CI = kjemisk ionisering (i relasjon til MS) CI = chemical ionization (relative to MS)

d = dag(er) d = day(s)

DCC = dicykloheksylkarbodiimid DCC = dicyclohexylcarbodiimide

DIBAL-H = di-isobutylaluminiumhydrid DIBAL-H = di-isobutyl aluminum hydride

DIPEA = diisopropyletylamin DIPEA = diisopropylethylamine

DMAP = 4-(AT,N-dimetylamino) pyridin DMAP = 4-(AT,N-dimethylamino)pyridine

DMF = dimetylformamid DMF = dimethylformamide

DMSO = dimetylsulfoksyd DMSO = dimethyl sulfoxide

DVT = dyp vene trombose DVT = deep vein thrombosis

EDC = l-(3-dimetylaminopropyl)-3-etylkarbodiimid-hydroklorid EDC = 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

e.e. = enantiomert overskudd av e.e. = enantiomeric excess of

Et = etyl Et = ethyl

ether = dietyleter ether = diethyl ether

EtOAc = etylacetat EtOAc = ethyl acetate

EtOH = etanol EtOH = ethanol

Et20 = dietyleter Et20 = diethyl ether

t = time(r) t = hour(s)

HATU = 0-(azabenzotriazol-l-yl)-iV,^V^V^V-tetrametyluronium heksafluorfosfat HATU = O-(azabenzotriazol-1-yl)-iV,^V^V^V-tetramethyluronium hexafluorophosphate

HBTU = [^,A^^^-tetrametyl-0-(benzotriazol-l-yl)uroniumheksafluorfosfat^ HC1 = saltsyre, hydrogenklorid gass eller hydrokloridsalt (avhengig av HBTU = [^,A^^^-tetramethyl-0-(benzotriazol-l-yl)uronium hexafluorophosphate^ HC1 = hydrochloric acid, hydrogen chloride gas or hydrochloride salt (depending on

sammenheng) context)

Heks = heksaner Hex = hexanes

HOAc = eddiksyre HOAc = acetic acid

HPLC = høy ytelse væskekromatografi HPLC = high performance liquid chromatography

LC = væskekromatografi LC = liquid chromatography

Me = metyl Me = methyl

MEM = metoksyetoksymetyl MEM = methoxyethoxymethyl

MeOH = metanol MeOH = methanol

min = minutt(er) min = minute(s)

MS = massespektroskopi MS = mass spectroscopy

MTBE = metyl terf-butyleter MTBE = methyl tert-butyl ether

NADH = nikotinamidadenindinukleotid, redusert form NADH = nicotinamide adenine dinucleotide, reduced form

NADPH = nikotinamidademndinukleotid fosfat, redusert form NADPH = nicotinamide adem dinucleotide phosphate, reduced form

NIH = "National Institute of Health "(US) NIH = "National Institute of Health" (US)

NIHU = "National Institute of Health" enheter NIHU = "National Institute of Health" units

NMR = kjernemagnetisk resonans NMR = nuclear magnetic resonance

OAc = acetat OAc = acetate

Pab = para-amidinobenzylamino Pab = para-amidinobenzylamino

H-Pab = parø-amidinobenzylamin H-Pab = para-amidinobenzylamine

Ph = fenyl Ph = phenyl

Pr = propyl Pr = propyl

Pro = (5)-prolinyl Pro = (5)-prolinyl

PyBOP = (benzotriazol-l-yloksy)tripyrrolidinofosfoniumheksafluorfosfat PyBOP = (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate

QF = tetrabutylammonium fluorid QF = tetrabutylammonium fluoride

RedAl = natrium bis(2-metoksyetoksy)aluminiumhydrid RedAl = sodium bis(2-methoxyethoxy)aluminum hydride

RPLC = revers fase høy ytelse væskekromatografi RPLC = reverse phase high performance liquid chromatography

rt/RT = romtemperatur rt/RT = room temperature

SOPs = standard drifts prosedyrer SOPs = standard operating procedures

TBTU = [^V^V^^-tetrametyl-O-^enzotriazol-l-yOuroniumtetrafluorborat] TE = trietylamin TBTU = [^V^V^^-tetramethyl-O-^enzotriazole-l-yOuronium tetrafluoroborate] TE = triethylamine

Teoc = 2-(trimetylsilyl)etoksykarbonyl Teoc = 2-(trimethylsilyl)ethoxycarbonyl

TEMPO = 2,2,6,6-tetrametyl-l-piperidinyloksy fri rest TEMPO = 2,2,6,6-tetramethyl-1-piperidinyloxy free residue

TFA = trifluoreddiksyre THF = tetrahydrofuran TFA = trifluoroacetic acid THF = tetrahydrofuran

THP = tetrahydropyranyl TLC = tynnskiktskromatografi TMSC1 = trimetylsilylklorid TMSCN = trimetylsilylcyanid UV = ultrafiolett THP = tetrahydropyranyl TLC = thin layer chromatography TMSC1 = trimethylsilyl chloride TMSCN = trimethylsilyl cyanide UV = ultraviolet

Z = benzyloksykarbonyl Z = benzyloxycarbonyl

Forstavelsene, s, i og t har deres vanlige betydninger: normal, sekundær, iso og tertiær. Forstavelsen c betyr cyklo. The prefixes, s, i and t have their usual meanings: normal, secondary, iso and tertiary. The prefix c means cyclo.

Claims (17)

1. Forbindelse, karakterisert ved at den er valgt fra Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe); Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OH) eller Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab, eller et farmasøytisk akseptabelt salt derav.1. Connection, characterized in that it is chosen from Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OMe); Ph(3-Cl)(5-OCHF2)-(/?)CH(OH)C(0)-Aze-Pab(OH) or Ph(3-Cl)(5-OCHF2)-(/?)CH( OH)C(0)-Aze-Pab, or a pharmaceutically acceptable salt thereof. 2. Forbindelse ifølge krav 1,karakterisert ved at den er Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(OMe) eller et farmasøytisk akseptebelt salt derav.2. Compound according to claim 1, characterized in that it is Ph(3-Cl)(5-OCHF2)-(Æ)CH(OH)C(0)-Aze-Pab(OMe) or a pharmaceutically acceptable salt thereof. 3. Farmasøytisk formulering, karakterisert ved at den omfatter hvilke som helst av forbindelsene ifølge krav 1 eller et farmasøytisk akseptabelt salt derav, blandet sammen med en farmasytisk akseptabel adjuvant, fortynningsmiddel eller bærer.3. Pharmaceutical formulation, characterized in that it comprises any of the compounds according to claim 1 or a pharmaceutically acceptable salt thereof, mixed together with a pharmaceutically acceptable adjuvant, diluent or carrier. 4. Farmasøytiskformuleringifølgekrav3,karakterisert ved at den omfatter forbindelsen ifølge krav 2, eller et farmasøytisk akseptabelt salt derav, blandet sammen med en farmasøytisk akseptabel adjuvant, fortynningsmiddel eller bærer.4. Pharmaceutical formulation according to claim 3, characterized in that it comprises the compound according to claim 2, or a pharmaceutically acceptable salt thereof, mixed together with a pharmaceutically acceptable adjuvant, diluent or carrier. 5. Farmasøytisk formulering ifølge krav 3 eller 4, karakterisert ved at adjuvanten, fortynningsmidlet og/eller bæreren gir opphav til en forbindelse med modifisert frigjøring.5. Pharmaceutical formulation according to claim 3 or 4, characterized in that the adjuvant, diluent and/or carrier give rise to a compound with modified release. 6. Farmasøytisk formulering ifølge krav 3 eller 4, karakterisert ved at den er tilpasset for oral administrering.6. Pharmaceutical formulation according to claim 3 or 4, characterized in that it is adapted for oral administration. 7. Farmasøytisk formulering ifølge krav 5 eller 6, karakterisert ved at den er i form av et geldannende matriksmodifisert-frigjøringssystem omfattende en hydrofil geldannende komponent og aktiv ingrediens.7. Pharmaceutical formulation according to claim 5 or 6, characterized in that it is in the form of a gel-forming matrix-modified release system comprising a hydrophilic gel-forming component and active ingredient. 8. Forbindelse ifølge krav 1 eller 2, karakterisert ved at den er for anvendelse som et farmasøytisk preparat.8. Compound according to claim 1 or 2, characterized in that it is for use as a pharmaceutical preparation. 9. Forbindelse ifølge krav 1 eller 2, karakterisert ved at den er for anvendelse ved behandling av en tilstand hvor inhibisjon av trombin er nødvendig.9. Compound according to claim 1 or 2, characterized in that it is for use in the treatment of a condition where inhibition of thrombin is necessary. 10. Forbindelse ifølge krav 1 eller 2, karakterisert ved at den er for anvendelse ved behandling av en tilstand hvor antikoagulantterapi er indikert.10. Compound according to claim 1 or 2, characterized in that it is for use in the treatment of a condition where anticoagulant therapy is indicated. 11. Forbindelse ifølge krav 1 eller 2, karakterisert ved at den er for anvendelse ved behandling av trombose.11. Compound according to claim 1 or 2, characterized in that it is for use in the treatment of thrombosis. 12. Forbindelse ifølge krav 1 eller 2, karakterisert ved at den er for anvendelse satt som et antikoaguleringsmiddel.12. Compound according to claim 1 or 2, characterized in that it is intended for use as an anticoagulant. 13. Anvendelse av en forbindelse ifølge krav 1 eller 2, som et aktivt ingrediens for fremstilling av et medikament for behandling av en tilstand hvor inhibisjon av trombin er nødvendig.13. Use of a compound according to claim 1 or 2, as an active ingredient for the preparation of a medicament for the treatment of a condition where inhibition of thrombin is necessary. 14. Anvendelse av en forbindelse ifølge krav 1 eller 2, som et aktivt ingrediens for fremstilling av et medikament for behandling av en tilstand hvor antikoaguleringsterapi er indikert.14. Use of a compound according to claim 1 or 2, as an active ingredient for the preparation of a medicament for the treatment of a condition where anticoagulation therapy is indicated. 15. Anvendelse ifølge krav 13 eller 14, hvor tilstanden er trombose.15. Use according to claim 13 or 14, where the condition is thrombosis. 16. Anvendelse ifølge krav 13 eller 14, hvor tilstanden er hyperkoagulabilitet i blod og/eller vev.16. Use according to claim 13 or 14, where the condition is hypercoagulability in blood and/or tissue. 17. Anvendelse av en forbindelse ifølge krav 1 eller 2 som et aktivt ingrediens for fremstilling av et antikoaguleringsmiddel.17. Use of a compound according to claim 1 or 2 as an active ingredient for the production of an anticoagulant.
NO20032465A 2000-12-01 2003-05-30 New mandelic acid derivatives and their use as well as pharmaceutical formulation NO325228B1 (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
SE0004458A SE0004458D0 (en) 2000-12-01 2000-12-01 Pharmaceutically useful compounds
SE0100965A SE0100965D0 (en) 2001-03-19 2001-03-19 Pharmaceutically useful compounds
SE0101239A SE0101239D0 (en) 2001-04-06 2001-04-06 Pharmaceutically useful compounds
SE0102921A SE0102921D0 (en) 2001-08-30 2001-08-30 Pharmaceutically useful compounds
PCT/SE2001/002657 WO2002044145A1 (en) 2000-12-01 2001-11-30 New mandelic acid derivatives and their use as throbin inhibitors

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